This document discusses using excipient combinations to reduce the viscosity of highly concentrated protein formulations for subcutaneous injection. High viscosity can make injections painful and even impossible. While single excipients sometimes lower viscosity, they may also destabilize proteins or require high concentrations that impact stability. The document finds that combining amino acids like ornithine or phenylalanine with anionic excipients can reduce viscosity more than single excipients, possibly through an over-additive effect. This allows using lower concentrations that better balance viscosity reduction and protein stability. Excipient performance also depends on formulation pH, as properties like a molecule's charge and hydrophobicity vary with pH. An excipient toolbox approach combining different excipients
The Viscosity Reduction Platform: Viscosity-reducing excipients for improveme...Merck Life Sciences
Protein viscosity is a major challenge in preparing highly concentrated protein formulations suitable for subcutaneous injection. Recently, the Viscosity Reduction Platform (VRP) was introduced and its technical key features and benefits for formulations were discussed. However, highly viscous solutions do not only pose a challenge when administering a drug to a patient, they can also impose technical limitations in the manufacturing process.
This white paper evaluates the effect of the excipients in the Viscosity Reduction Platform on ultrafiltration processes used to produce a highly concentrated formulation of a monoclonal antibody (mAb). Two filtration methods are demonstrated in this work.
Find more information about the Viscosity Reduction Platform on our website: https://www.sigmaaldrich.com/products/pharma-and-biopharma-manufacturing/formulation/viscosity-reduction-platform
The Viscosity Reduction Platform: Viscosity Reducing Excipients for Protein F...Merck Life Sciences
Protein viscosity is one of the major obstacles in preparing highly concentrated protein formulations suitable for subcutaneous injection.
This whitepaper examines how combining an amino acid with a second viscosity-reducing excipient circumvents adverse effects on protein stability and improves viscosity-reducing capacity.
To find more information about the Viscosity Reduction Platform, please visit our website: https://sigmaaldrich.com/products/pharma-and-biopharma-manufacturing/formulation/viscosity-reduction-platform
The Viscosity Reduction Platform: Viscosity-reducing excipients for improveme...MilliporeSigma
Protein viscosity is a major challenge in preparing highly concentrated protein formulations suitable for subcutaneous injection. Recently, the Viscosity Reduction Platform (VRP) was introduced and its technical key features and benefits for formulations were discussed. However, highly viscous solutions do not only pose a challenge when administering a drug to a patient, they can also impose technical limitations in the manufacturing process.
This white paper evaluates the effect of the excipients in the Viscosity Reduction Platform on ultrafiltration processes used to produce a highly concentrated formulation of a monoclonal antibody (mAb). Two filtration methods are demonstrated in this work.
Find more information about the Viscosity Reduction Platform on our website: https://www.sigmaaldrich.com/products/pharma-and-biopharma-manufacturing/formulation/viscosity-reduction-platform
The Viscosity Reduction Platform: Viscosity Reducing Excipients for Protein F...MilliporeSigma
Protein viscosity is one of the major obstacles in preparing highly concentrated protein formulations suitable for subcutaneous injection.
This whitepaper examines how combining an amino acid with a second viscosity-reducing excipient circumvents adverse effects on protein stability and improves viscosity-reducing capacity.
To find more information about the Viscosity Reduction Platform, please visit our website: https://sigmaaldrich.com/products/pharma-and-biopharma-manufacturing/formulation/viscosity-reduction-platform
The Viscosity Reduction Platform: Viscosity-reducing excipients for improveme...Merck Life Sciences
Protein viscosity is a major challenge in preparing highly concentrated protein formulations suitable for subcutaneous injection. Recently, the Viscosity Reduction Platform (VRP) was introduced and its technical key features and benefits for formulations were discussed. However, highly viscous solutions do not only pose a challenge when administering a drug to a patient, they can also impose technical limitations in the manufacturing process.
This white paper evaluates the effect of the excipients in the Viscosity Reduction Platform on ultrafiltration processes used to produce a highly concentrated formulation of a monoclonal antibody (mAb). Two filtration methods are demonstrated in this work.
Find more information about the Viscosity Reduction Platform on our website: https://www.sigmaaldrich.com/products/pharma-and-biopharma-manufacturing/formulation/viscosity-reduction-platform
The Viscosity Reduction Platform: Viscosity Reducing Excipients for Protein F...Merck Life Sciences
Protein viscosity is one of the major obstacles in preparing highly concentrated protein formulations suitable for subcutaneous injection.
This whitepaper examines how combining an amino acid with a second viscosity-reducing excipient circumvents adverse effects on protein stability and improves viscosity-reducing capacity.
To find more information about the Viscosity Reduction Platform, please visit our website: https://sigmaaldrich.com/products/pharma-and-biopharma-manufacturing/formulation/viscosity-reduction-platform
The Viscosity Reduction Platform: Viscosity-reducing excipients for improveme...MilliporeSigma
Protein viscosity is a major challenge in preparing highly concentrated protein formulations suitable for subcutaneous injection. Recently, the Viscosity Reduction Platform (VRP) was introduced and its technical key features and benefits for formulations were discussed. However, highly viscous solutions do not only pose a challenge when administering a drug to a patient, they can also impose technical limitations in the manufacturing process.
This white paper evaluates the effect of the excipients in the Viscosity Reduction Platform on ultrafiltration processes used to produce a highly concentrated formulation of a monoclonal antibody (mAb). Two filtration methods are demonstrated in this work.
Find more information about the Viscosity Reduction Platform on our website: https://www.sigmaaldrich.com/products/pharma-and-biopharma-manufacturing/formulation/viscosity-reduction-platform
The Viscosity Reduction Platform: Viscosity Reducing Excipients for Protein F...MilliporeSigma
Protein viscosity is one of the major obstacles in preparing highly concentrated protein formulations suitable for subcutaneous injection.
This whitepaper examines how combining an amino acid with a second viscosity-reducing excipient circumvents adverse effects on protein stability and improves viscosity-reducing capacity.
To find more information about the Viscosity Reduction Platform, please visit our website: https://sigmaaldrich.com/products/pharma-and-biopharma-manufacturing/formulation/viscosity-reduction-platform
Improving Downstream Processing: Application of Excipients in DSPMilliporeSigma
This study investigates the benefits of adding excipients during downstream processing on protein stability, chromatographic performance and viral inactivation.
Mucilage of basil seed can be employed as a potential ingredient in suspensions, emulsions, gels and tablets especially as viscosity enhancing agents, thickening agent, emulsifier or gelling agent and release retardant because of its good hydrophilic nature, physical stability, barrier properties, efficient control of release profile, extrudability and good spreadability.
Extensive characterisation of natural polymer in dosage form development for subsequent commercialisation has given rise to a new term “Naturapolyceutics”.
Use of Excipients in Downstream Processing to Improve Protein PurificationMerck Life Sciences
Excipients are used to improve the stability of protein-based therapeutics by protecting the protein against a range of stress conditions such as temperature changes, pH changes, or agitation. Similar stresses are applied to proteins during downstream purification. Shifts in pH during Protein A chromatography, subsequent incubations at low pH for virus inactivation, and changes in conductivity in ion exchange chromatography can lead to aggregation, fragmentation, or other chemical modifications of the therapeutic protein. Given the potential impact on the protein’s structural integrity, there is a need for approaches to reduce the risk presented by the conditions during downstream processing. For example, integration of a solution to prevent aggregation of proteins would be a more efficient strategy than implementing steps to remove multimeric forms.
This white paper highlights the results from a recent paper by Stange et. al., in which protein stabilizing excipients such as polyols, sugars, and polyethylene glycol (PEG4000) were used as buffer system additives. Effect of the excipients on elution patterns, stabilization of the monomer antibody, host-cell protein removal, virus inactivation rates and binding capacity of cation exchange chromatography were explored.
Use of Excipients in Downstream Processing to Improve Protein PurificationMilliporeSigma
Excipients are used to improve the stability of protein-based therapeutics by protecting the protein against a range of stress conditions such as temperature changes, pH changes, or agitation. Similar stresses are applied to proteins during downstream purification. Shifts in pH during Protein A chromatography, subsequent incubations at low pH for virus inactivation, and changes in conductivity in ion exchange chromatography can lead to aggregation, fragmentation, or other chemical modifications of the therapeutic protein. Given the potential impact on the protein’s structural integrity, there is a need for approaches to reduce the risk presented by the conditions during downstream processing. For example, integration of a solution to prevent aggregation of proteins would be a more efficient strategy than implementing steps to remove multimeric forms.
This white paper highlights the results from a recent paper by Stange et. al., in which protein stabilizing excipients such as polyols, sugars, and polyethylene glycol (PEG4000) were used as buffer system additives. Effect of the excipients on elution patterns, stabilization of the monomer antibody, host-cell protein removal, virus inactivation rates and binding capacity of cation exchange chromatography were explored.
The Viscosity Reduction Platform: Enabling Subcutaneous (subQ) DeliveryMilliporeSigma
Watch the presentation of this webinar here: https://bit.ly/3FKGUH6
At the high protein concentrations required for subcutaneous administration, protein formulations often become highly viscous. In this webinar, Dr. Tobias Rosenkranz will introduce a new approach that combines different excipients to reduce viscosity and discuss synergistic effects.
Subcutaneous (subQ) administration can improve patient convenience and reduce healthcare costs by avoiding the need for hospitalization. Yet in some cases, high protein concentrations make formulations far more viscous, prohibiting this route of administration. While viscosity can normally be reduced by using certain excipients, merely adding more and more of a single excipient may not bring sufficient improvement and can even compromise protein stability. This webinar will introduce an excipient platform that makes it possible to combine excipients in ways that can reduce protein viscosity to a greater extent.
In this webinar, you will learn about:
• Challenges arising from high concentrated protein formulations
• The viscosity reduction platform: a portfolio of excipients to manage protein viscosity
• The impact of viscosity reducing excipients on protein stability
• The impact of protein viscosity on syringability
Presented by: Tobias Rosenkranz, Ph.D.,
Senior Manager, Biomolecule Formulation R&D
The Viscosity Reduction Platform: Enabling Subcutaneous (subQ) DeliveryMerck Life Sciences
Watch the presentation of this webinar here: https://bit.ly/3FKGUH6
At the high protein concentrations required for subcutaneous administration, protein formulations often become highly viscous. In this webinar, Dr. Tobias Rosenkranz will introduce a new approach that combines different excipients to reduce viscosity and discuss synergistic effects.
Subcutaneous (subQ) administration can improve patient convenience and reduce healthcare costs by avoiding the need for hospitalization. Yet in some cases, high protein concentrations make formulations far more viscous, prohibiting this route of administration. While viscosity can normally be reduced by using certain excipients, merely adding more and more of a single excipient may not bring sufficient improvement and can even compromise protein stability. This webinar will introduce an excipient platform that makes it possible to combine excipients in ways that can reduce protein viscosity to a greater extent.
In this webinar, you will learn about:
• Challenges arising from high concentrated protein formulations
• The viscosity reduction platform: a portfolio of excipients to manage protein viscosity
• The impact of viscosity reducing excipients on protein stability
• The impact of protein viscosity on syringability
Presented by: Tobias Rosenkranz, Ph.D., Senior Manager, Biomolecule Formulation R&D
Exploring the protein stabilizing capability of surfactants against agitation...MilliporeSigma
Agitation of therapeutic protein solutions during manufacturing, shipping and handling is one of the major initiators for protein aggregation and particle formation during the life history of a protein drug. Adsorption of protein molecules to liquid-air interfaces leads to the formation of highly concentrated protein surface films. The rupture of these protein films due to various mechanical processes can then result in the appearance of protein aggregates and particles in the bulk solution phase.
One technique to stabilize proteins against stress induced by liquid-air interfaces is the use of non-ionic surfactants. About 91% of antibody formulations commercially available in 2021 contained a surfactant. Polysorbate 20 and 80, composed of a hydrophilic polyoxyethylene sorbitan and hydrophobic fatty acid esters, made up the largest part being employed in 87% of said formulations.
Despite their frequent use in parenteral drug products, concerns have been raised for decades about the application of polysorbates as surfactants in biopharmaceutical formulations. Autoxidation of polysorbate, caused by residual peroxides in polysorbates, can damage the proteins and can further drive the oxidative degradation of polysorbate. Chemical and enzymatic hydrolysis of polysorbate may lead to the formation of free fatty acid particles, which may become visible; and both mechanisms eventually lead to the reduction in polysorbate concentration. Therefore, the purpose of the current study was to compare various molecules for their capabilities to reduced agitation-induced protein aggregation and particle formation; and furthermore, investigate their underlying protein stabilizing mechanisms.
Protecting Protein Stability with a Novel Grade of SucroseMerck Life Sciences
Sucrose is one of the most widely used stabilizers in marketed drug products, ensuring the chemical and physical stability of the therapeutic protein. A challenge with the use of sucrose as an excipient is that nanoparticle impurities (NPI) can originate from the raw materials or be introduced during production.
In this whitepaper, you can find information on NPIs found in commercially available sucrose, their origin and impact on protein stability; and on a novel sucrose purification process designed to minimize the presence of NPIs.
To find more information about this novel grade of sucrose, please visit our website: https://www.sigmaaldrich.com/product/mm/103789
And to learn more about protein stabilization of biomolecules in general, please follow this link: https://www.sigmaaldrich.com/products/pharma-and-biopharma-manufacturing/formulation/protein-stabilizers
Exploring the protein stabilizing capability of surfactants against agitation...Merck Life Sciences
Agitation of therapeutic protein solutions during manufacturing, shipping and handling is one of the major initiators for protein aggregation and particle formation during the life history of a protein drug. Adsorption of protein molecules to liquid-air interfaces leads to the formation of highly concentrated protein surface films. The rupture of these protein films due to various mechanical processes can then result in the appearance of protein aggregates and particles in the bulk solution phase.
One technique to stabilize proteins against stress induced by liquid-air interfaces is the use of non-ionic surfactants. About 91% of antibody formulations commercially available in 2021 contained a surfactant. Polysorbate 20 and 80, composed of a hydrophilic polyoxyethylene sorbitan and hydrophobic fatty acid esters, made up the largest part being employed in 87% of said formulations.
Despite their frequent use in parenteral drug products, concerns have been raised for decades about the application of polysorbates as surfactants in biopharmaceutical formulations. Autoxidation of polysorbate, caused by residual peroxides in polysorbates, can damage the proteins and can further drive the oxidative degradation of polysorbate. Chemical and enzymatic hydrolysis of polysorbate may lead to the formation of free fatty acid particles, which may become visible; and both mechanisms eventually lead to the reduction in polysorbate concentration. Therefore, the purpose of the current study was to compare various molecules for their capabilities to reduced agitation-induced protein aggregation and particle formation; and furthermore, investigate their underlying protein stabilizing mechanisms.
Improving Downstream Processing: Application of Excipients in DSPMerck Life Sciences
Webinar summary:
This webinar will showcase the beneficial potential of using excipients during downstream processing of monoclonal antibodies.
Learning points:
In this webinar, you will see:
* An innovative excipient screening approach simulating low pH stress conditions during protein A chromatography and virus inactivation
* How the application of excipients in buffer systems can significantly improve protein stability and chromatographic performance
Abstract:
Key aspects during downstream purification of biopharmaceutical drugs are purity and process yield. Therefore, the downstream process needs to be designed in a way that the final product which will eventually end up in the patient entails low levels of product- and process related impurities (e.g. high molecular weight aggregates) as well as process related contaminants (e.g. host cell protein levels). In addition to this, the process must be capable of clearing and inactivating viruses to ensure product safety. In this webinar, we will explore the benefits of adding excipients during downstream processing on protein stability, chromatographic performance and viral inactivation.
Improving Downstream Processing: Application of Excipients in DSPMilliporeSigma
Webinar summary:
This webinar will showcase the beneficial potential of using excipients during downstream processing of monoclonal antibodies.
Learning points:
In this webinar, you will see:
* An innovative excipient screening approach simulating low pH stress conditions during protein A chromatography and virus inactivation
* How the application of excipients in buffer systems can significantly improve protein stability and chromatographic performance
Abstract:
Key aspects during downstream purification of biopharmaceutical drugs are purity and process yield. Therefore, the downstream process needs to be designed in a way that the final product which will eventually end up in the patient entails low levels of product- and process related impurities (e.g. high molecular weight aggregates) as well as process related contaminants (e.g. host cell protein levels). In addition to this, the process must be capable of clearing and inactivating viruses to ensure product safety. In this webinar, we will explore the benefits of adding excipients during downstream processing on protein stability, chromatographic performance and viral inactivation.
Effect of solvents on formation of disulphide bond in peptides: A comparative...inventionjournals
A novel approach to the formation of disulphide bonds in peptides is developed by doing a series of
experiments to evaluate effect of solvents on the disulphide bond formation in peptides in terms of time required,
recovery, ease of processing during reaction working up and use of solvents. For evaluating the effect,
experiments were setup using Water:Acetonitrile, Water:Methanol and Water:Ethanol for disulphide bond
formation of Desmopressin and the results were compared with the aerial oxidation conducted in water. For
each solvent, a set of oxidation reactions was studied at 0.5 mg/ml, 1 mg/ml and 5 mg/ml concentration. The
acetonitrile water combination at concentration of either 0.5mg/ml or 1 mg/ml proved to deliver the best result
in terms of purity, yield and time required to complete reaction.
Effect of solvents on formation of disulphide bond in peptides: A comparativ...inventionjournals
International Journal of Pharmaceutical Science Invention (IJPSI) is an international journal intended for professionals and researchers in all fields of Pahrmaceutical Science. IJPSI publishes research articles and reviews within the whole field Pharmacy and Pharmaceutical Science, new teaching methods, assessment, validation and the impact of new technologies and it will continue to provide information on the latest trends and developments in this ever-expanding subject. The publications of papers are selected through double peer reviewed to ensure originality, relevance, and readability. The articles published in our journal can be accessed online.
Characterization of monoclonal antibodies and Antibody drug conjugates by Sur...MilliporeSigma
Watch the presentation of this webinar: https://bit.ly/3Pjpjvr
Highlights of this webinar:
- Surface plasmon resonance as a powerful tool for biologic characterization including mAbs and ADCs.
- SPR allows rapid binding analysis in real time without using labels for SARS-CoV-2 receptor binding domain mutations.
- Kinetic data is indicative of possible neutralizing activity allowed assessment of neutralizing ability of therapeutic monoclonal antibodies.
- The application can provide preliminarily efficacy information and facilitated mAbs/ACDs candidate selection process
Detailed description:
Characterization of therapeutic monoclonal antibodies (mAbs) or Antibody drug conjugates (ADCs) is challenging due to their ability to bind to a variety of proteins via their Fc and Fab domains, giving rise to diverse biological functions associated with each domain. The Fc domain of mAbs interacts with Fc receptors with varying affinities, which can influence biological processes such as Complement-dependent cytotoxicity (CDC) and Antibody-dependent cellular cytotoxicity (ADCC), transcytosis, phagocytosis, and/or serum half-life.
An important characteristic of an antibody is its Fc effector function. Antibodies can be engineered to obtain desired binding of the Fc region to Fc receptors expressed on effector cells. Hence, it is crucial to evaluate the binding interaction of mAbs/ADC with Fc receptors in the early phase of drug development to understand the potential biological activity of the product in vivo.
Surface Plasmon Resonance (SPR) is a powerful technique to establish binding kinetics in real-time, label free, and high sensitivity with low sample consumption. Along with target antigen binding, it is crucial to evaluate the binding interaction of antibodies and ADCs with Fc receptors. Our SPR case studies investigated the impact on binding kinetics of ADCs with different linkers and the binding interactions of SARS-CoV-2 spike protein variants and evaluated the neutralizing ability of therapeutic mAbs. SPR characterisation can be facilitated in all stages of the product life cycle to ensure the quality and safety of mAbs and ADCs.
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Improving Downstream Processing: Application of Excipients in DSPMilliporeSigma
This study investigates the benefits of adding excipients during downstream processing on protein stability, chromatographic performance and viral inactivation.
Mucilage of basil seed can be employed as a potential ingredient in suspensions, emulsions, gels and tablets especially as viscosity enhancing agents, thickening agent, emulsifier or gelling agent and release retardant because of its good hydrophilic nature, physical stability, barrier properties, efficient control of release profile, extrudability and good spreadability.
Extensive characterisation of natural polymer in dosage form development for subsequent commercialisation has given rise to a new term “Naturapolyceutics”.
Use of Excipients in Downstream Processing to Improve Protein PurificationMerck Life Sciences
Excipients are used to improve the stability of protein-based therapeutics by protecting the protein against a range of stress conditions such as temperature changes, pH changes, or agitation. Similar stresses are applied to proteins during downstream purification. Shifts in pH during Protein A chromatography, subsequent incubations at low pH for virus inactivation, and changes in conductivity in ion exchange chromatography can lead to aggregation, fragmentation, or other chemical modifications of the therapeutic protein. Given the potential impact on the protein’s structural integrity, there is a need for approaches to reduce the risk presented by the conditions during downstream processing. For example, integration of a solution to prevent aggregation of proteins would be a more efficient strategy than implementing steps to remove multimeric forms.
This white paper highlights the results from a recent paper by Stange et. al., in which protein stabilizing excipients such as polyols, sugars, and polyethylene glycol (PEG4000) were used as buffer system additives. Effect of the excipients on elution patterns, stabilization of the monomer antibody, host-cell protein removal, virus inactivation rates and binding capacity of cation exchange chromatography were explored.
Use of Excipients in Downstream Processing to Improve Protein PurificationMilliporeSigma
Excipients are used to improve the stability of protein-based therapeutics by protecting the protein against a range of stress conditions such as temperature changes, pH changes, or agitation. Similar stresses are applied to proteins during downstream purification. Shifts in pH during Protein A chromatography, subsequent incubations at low pH for virus inactivation, and changes in conductivity in ion exchange chromatography can lead to aggregation, fragmentation, or other chemical modifications of the therapeutic protein. Given the potential impact on the protein’s structural integrity, there is a need for approaches to reduce the risk presented by the conditions during downstream processing. For example, integration of a solution to prevent aggregation of proteins would be a more efficient strategy than implementing steps to remove multimeric forms.
This white paper highlights the results from a recent paper by Stange et. al., in which protein stabilizing excipients such as polyols, sugars, and polyethylene glycol (PEG4000) were used as buffer system additives. Effect of the excipients on elution patterns, stabilization of the monomer antibody, host-cell protein removal, virus inactivation rates and binding capacity of cation exchange chromatography were explored.
The Viscosity Reduction Platform: Enabling Subcutaneous (subQ) DeliveryMilliporeSigma
Watch the presentation of this webinar here: https://bit.ly/3FKGUH6
At the high protein concentrations required for subcutaneous administration, protein formulations often become highly viscous. In this webinar, Dr. Tobias Rosenkranz will introduce a new approach that combines different excipients to reduce viscosity and discuss synergistic effects.
Subcutaneous (subQ) administration can improve patient convenience and reduce healthcare costs by avoiding the need for hospitalization. Yet in some cases, high protein concentrations make formulations far more viscous, prohibiting this route of administration. While viscosity can normally be reduced by using certain excipients, merely adding more and more of a single excipient may not bring sufficient improvement and can even compromise protein stability. This webinar will introduce an excipient platform that makes it possible to combine excipients in ways that can reduce protein viscosity to a greater extent.
In this webinar, you will learn about:
• Challenges arising from high concentrated protein formulations
• The viscosity reduction platform: a portfolio of excipients to manage protein viscosity
• The impact of viscosity reducing excipients on protein stability
• The impact of protein viscosity on syringability
Presented by: Tobias Rosenkranz, Ph.D.,
Senior Manager, Biomolecule Formulation R&D
The Viscosity Reduction Platform: Enabling Subcutaneous (subQ) DeliveryMerck Life Sciences
Watch the presentation of this webinar here: https://bit.ly/3FKGUH6
At the high protein concentrations required for subcutaneous administration, protein formulations often become highly viscous. In this webinar, Dr. Tobias Rosenkranz will introduce a new approach that combines different excipients to reduce viscosity and discuss synergistic effects.
Subcutaneous (subQ) administration can improve patient convenience and reduce healthcare costs by avoiding the need for hospitalization. Yet in some cases, high protein concentrations make formulations far more viscous, prohibiting this route of administration. While viscosity can normally be reduced by using certain excipients, merely adding more and more of a single excipient may not bring sufficient improvement and can even compromise protein stability. This webinar will introduce an excipient platform that makes it possible to combine excipients in ways that can reduce protein viscosity to a greater extent.
In this webinar, you will learn about:
• Challenges arising from high concentrated protein formulations
• The viscosity reduction platform: a portfolio of excipients to manage protein viscosity
• The impact of viscosity reducing excipients on protein stability
• The impact of protein viscosity on syringability
Presented by: Tobias Rosenkranz, Ph.D., Senior Manager, Biomolecule Formulation R&D
Exploring the protein stabilizing capability of surfactants against agitation...MilliporeSigma
Agitation of therapeutic protein solutions during manufacturing, shipping and handling is one of the major initiators for protein aggregation and particle formation during the life history of a protein drug. Adsorption of protein molecules to liquid-air interfaces leads to the formation of highly concentrated protein surface films. The rupture of these protein films due to various mechanical processes can then result in the appearance of protein aggregates and particles in the bulk solution phase.
One technique to stabilize proteins against stress induced by liquid-air interfaces is the use of non-ionic surfactants. About 91% of antibody formulations commercially available in 2021 contained a surfactant. Polysorbate 20 and 80, composed of a hydrophilic polyoxyethylene sorbitan and hydrophobic fatty acid esters, made up the largest part being employed in 87% of said formulations.
Despite their frequent use in parenteral drug products, concerns have been raised for decades about the application of polysorbates as surfactants in biopharmaceutical formulations. Autoxidation of polysorbate, caused by residual peroxides in polysorbates, can damage the proteins and can further drive the oxidative degradation of polysorbate. Chemical and enzymatic hydrolysis of polysorbate may lead to the formation of free fatty acid particles, which may become visible; and both mechanisms eventually lead to the reduction in polysorbate concentration. Therefore, the purpose of the current study was to compare various molecules for their capabilities to reduced agitation-induced protein aggregation and particle formation; and furthermore, investigate their underlying protein stabilizing mechanisms.
Protecting Protein Stability with a Novel Grade of SucroseMerck Life Sciences
Sucrose is one of the most widely used stabilizers in marketed drug products, ensuring the chemical and physical stability of the therapeutic protein. A challenge with the use of sucrose as an excipient is that nanoparticle impurities (NPI) can originate from the raw materials or be introduced during production.
In this whitepaper, you can find information on NPIs found in commercially available sucrose, their origin and impact on protein stability; and on a novel sucrose purification process designed to minimize the presence of NPIs.
To find more information about this novel grade of sucrose, please visit our website: https://www.sigmaaldrich.com/product/mm/103789
And to learn more about protein stabilization of biomolecules in general, please follow this link: https://www.sigmaaldrich.com/products/pharma-and-biopharma-manufacturing/formulation/protein-stabilizers
Exploring the protein stabilizing capability of surfactants against agitation...Merck Life Sciences
Agitation of therapeutic protein solutions during manufacturing, shipping and handling is one of the major initiators for protein aggregation and particle formation during the life history of a protein drug. Adsorption of protein molecules to liquid-air interfaces leads to the formation of highly concentrated protein surface films. The rupture of these protein films due to various mechanical processes can then result in the appearance of protein aggregates and particles in the bulk solution phase.
One technique to stabilize proteins against stress induced by liquid-air interfaces is the use of non-ionic surfactants. About 91% of antibody formulations commercially available in 2021 contained a surfactant. Polysorbate 20 and 80, composed of a hydrophilic polyoxyethylene sorbitan and hydrophobic fatty acid esters, made up the largest part being employed in 87% of said formulations.
Despite their frequent use in parenteral drug products, concerns have been raised for decades about the application of polysorbates as surfactants in biopharmaceutical formulations. Autoxidation of polysorbate, caused by residual peroxides in polysorbates, can damage the proteins and can further drive the oxidative degradation of polysorbate. Chemical and enzymatic hydrolysis of polysorbate may lead to the formation of free fatty acid particles, which may become visible; and both mechanisms eventually lead to the reduction in polysorbate concentration. Therefore, the purpose of the current study was to compare various molecules for their capabilities to reduced agitation-induced protein aggregation and particle formation; and furthermore, investigate their underlying protein stabilizing mechanisms.
Improving Downstream Processing: Application of Excipients in DSPMerck Life Sciences
Webinar summary:
This webinar will showcase the beneficial potential of using excipients during downstream processing of monoclonal antibodies.
Learning points:
In this webinar, you will see:
* An innovative excipient screening approach simulating low pH stress conditions during protein A chromatography and virus inactivation
* How the application of excipients in buffer systems can significantly improve protein stability and chromatographic performance
Abstract:
Key aspects during downstream purification of biopharmaceutical drugs are purity and process yield. Therefore, the downstream process needs to be designed in a way that the final product which will eventually end up in the patient entails low levels of product- and process related impurities (e.g. high molecular weight aggregates) as well as process related contaminants (e.g. host cell protein levels). In addition to this, the process must be capable of clearing and inactivating viruses to ensure product safety. In this webinar, we will explore the benefits of adding excipients during downstream processing on protein stability, chromatographic performance and viral inactivation.
Improving Downstream Processing: Application of Excipients in DSPMilliporeSigma
Webinar summary:
This webinar will showcase the beneficial potential of using excipients during downstream processing of monoclonal antibodies.
Learning points:
In this webinar, you will see:
* An innovative excipient screening approach simulating low pH stress conditions during protein A chromatography and virus inactivation
* How the application of excipients in buffer systems can significantly improve protein stability and chromatographic performance
Abstract:
Key aspects during downstream purification of biopharmaceutical drugs are purity and process yield. Therefore, the downstream process needs to be designed in a way that the final product which will eventually end up in the patient entails low levels of product- and process related impurities (e.g. high molecular weight aggregates) as well as process related contaminants (e.g. host cell protein levels). In addition to this, the process must be capable of clearing and inactivating viruses to ensure product safety. In this webinar, we will explore the benefits of adding excipients during downstream processing on protein stability, chromatographic performance and viral inactivation.
Effect of solvents on formation of disulphide bond in peptides: A comparative...inventionjournals
A novel approach to the formation of disulphide bonds in peptides is developed by doing a series of
experiments to evaluate effect of solvents on the disulphide bond formation in peptides in terms of time required,
recovery, ease of processing during reaction working up and use of solvents. For evaluating the effect,
experiments were setup using Water:Acetonitrile, Water:Methanol and Water:Ethanol for disulphide bond
formation of Desmopressin and the results were compared with the aerial oxidation conducted in water. For
each solvent, a set of oxidation reactions was studied at 0.5 mg/ml, 1 mg/ml and 5 mg/ml concentration. The
acetonitrile water combination at concentration of either 0.5mg/ml or 1 mg/ml proved to deliver the best result
in terms of purity, yield and time required to complete reaction.
Effect of solvents on formation of disulphide bond in peptides: A comparativ...inventionjournals
International Journal of Pharmaceutical Science Invention (IJPSI) is an international journal intended for professionals and researchers in all fields of Pahrmaceutical Science. IJPSI publishes research articles and reviews within the whole field Pharmacy and Pharmaceutical Science, new teaching methods, assessment, validation and the impact of new technologies and it will continue to provide information on the latest trends and developments in this ever-expanding subject. The publications of papers are selected through double peer reviewed to ensure originality, relevance, and readability. The articles published in our journal can be accessed online.
Similar to The Viscosity Reduction Platform: Viscosity-Reducing Excipients for Protein Formulation (20)
Characterization of monoclonal antibodies and Antibody drug conjugates by Sur...MilliporeSigma
Watch the presentation of this webinar: https://bit.ly/3Pjpjvr
Highlights of this webinar:
- Surface plasmon resonance as a powerful tool for biologic characterization including mAbs and ADCs.
- SPR allows rapid binding analysis in real time without using labels for SARS-CoV-2 receptor binding domain mutations.
- Kinetic data is indicative of possible neutralizing activity allowed assessment of neutralizing ability of therapeutic monoclonal antibodies.
- The application can provide preliminarily efficacy information and facilitated mAbs/ACDs candidate selection process
Detailed description:
Characterization of therapeutic monoclonal antibodies (mAbs) or Antibody drug conjugates (ADCs) is challenging due to their ability to bind to a variety of proteins via their Fc and Fab domains, giving rise to diverse biological functions associated with each domain. The Fc domain of mAbs interacts with Fc receptors with varying affinities, which can influence biological processes such as Complement-dependent cytotoxicity (CDC) and Antibody-dependent cellular cytotoxicity (ADCC), transcytosis, phagocytosis, and/or serum half-life.
An important characteristic of an antibody is its Fc effector function. Antibodies can be engineered to obtain desired binding of the Fc region to Fc receptors expressed on effector cells. Hence, it is crucial to evaluate the binding interaction of mAbs/ADC with Fc receptors in the early phase of drug development to understand the potential biological activity of the product in vivo.
Surface Plasmon Resonance (SPR) is a powerful technique to establish binding kinetics in real-time, label free, and high sensitivity with low sample consumption. Along with target antigen binding, it is crucial to evaluate the binding interaction of antibodies and ADCs with Fc receptors. Our SPR case studies investigated the impact on binding kinetics of ADCs with different linkers and the binding interactions of SARS-CoV-2 spike protein variants and evaluated the neutralizing ability of therapeutic mAbs. SPR characterisation can be facilitated in all stages of the product life cycle to ensure the quality and safety of mAbs and ADCs.
The Role of BioPhorum Extractables Data in the Effective Adoption of Single-U...MilliporeSigma
Regulatory expectation does require patient safety evaluations with supporting data for manufacturing components that directly come into contact with drug manufacturing process streams. Readily available extractables data can help manufacturers using singleuse technology to accelerate product qualifications, risk assessments and process optimization
This white paper guides you on how to save time and resources with supplier-provided single-use system extractables data and gives you an overview about the overall strategy for Extractables & Leachables. At the end you will find a case study.
Find more information about filters and single-use components on our website: https://www.sigmaaldrich.com/DE/en/services/product-services/emprove-program/emprove-filter-and-single-use-component-portfolio
The Future of Pharma- and Biopharmaceutical AuditsMilliporeSigma
Watch the recording of this presentation here: https://bit.ly/3zTOpe4
Detailed description:
SARS-CoV-2 showed us that technology supports us during our inspection activity even if on-site visits are not possible. Travel restrictions of various kinds will remain a risk in the future. The use of new technologies has shown that inspections and audits can be carried out despite these restrictions. We will focus on what possibilities the new technologies offer and take a look at the future of inspections and audits.
In this webinar, you will learn:
• Regulatory overview of remote audits
• The technologies needed to support the audit process
• What types of inspections are possible with the use of these technologies
• How audits may look in the future
Presented by:
Daniel Buescher, Product Manager - Digital Solutions
Moving your Gene Therapy from R&D to IND: How to navigate the Regulatory Land...MilliporeSigma
Watch the recording of this presentation here: https://bit.ly/3SqOsoP
Novel therapies, including cell and gene therapies, continue to be central to innovation in healthcare and represent the fastest growing area of therapeutic medicine. As a consequence, the number of gene therapies undergoing clinical trials has increased significantly in the last five years.
Manufacturing processes for these novel therapeutics are very complex with a high risk of contamination. Regulatory agencies world-wide have responded by issuing guidance to outline their expectations for development and manufacture of cell and gene therapies. Currently, regulatory guidance is not harmonized globally and can often lead to confusion within industry and increased risk of non-compliance.
In this webinar, we'll answer:
• Which regulatory guidelines do you need to comply for your INDs?
• When do you start implementing GMPs and validated assays?
• How do you get your QC testing strategy ‘right the first time’?
• How do you ensure testing is not your rate limiting step for the IND submission?
Presented by:
Manjula Aysola, Senior Regulatory Consultant
Dr. Alison Armstrong, Sr. Director, Technical and Scientific Solutions
Identity testing by NGS as a means of risk mitigation for viral gene therapiesMilliporeSigma
Watch the presentation of this webinar here: https://bit.ly/3RijkHC
Detailed description:
Imagine you’ve just completed a manufacturing run for your viral vector. Identity testing is performed to confirm the vector sequence. But when the results come back the data reveals unexpected sequence variants! With an appropriate risk mitigation testing strategy, this situation can be prevented.
The situation described above is not hypothetical, and happens more that you think, costing valuable time and resources.
Investigatory testing has shown that sequence variants present in starting materials (e.g. plasmids) are likely to make their way to the final product. Adequate identification of low-level variants with an appropriately sensitive method is critical in ensuring the quality of the final product. A risk-based testing strategy, in the context of identity, for viral vector manufacturing will be presented, focusing on key testing points. NGS assays for identity and variant detection will be highlighted due to their extremely sensitive nature compared to traditional approaches.
In this webinar, we'll explore:
• Regulatory requirements for identity testing
• NGS applications for identity testing as compared to traditional methods
• A case study on the impact of not establishing a proper risk-based testing strategy
Presented by: Bradley Hasson, Director of Lab Operations for NGS Services
Latest advancements of melt based 3D printing technologies for oral drug deli...MilliporeSigma
Watch the presentation of this webinar here: https://bit.ly/3A2WcH4
The application of polymer excipients in 3D printing manufacturing is usually limited due to the concerns of filament strength, high processing temperature and large scale manufacturing.
Latest technology developments are targeting a direct melt deposition to simplify the process and enable a constant and efficient process. Two different processing approaches will be presented:
The advanced melt drop deposition, where individual three dimensional geometries can be created by depostition of polymer droplets and the MED® 3D printing technology which allows by precise layer-by-layer deposition to produce objects with well-designed geometric structures.
In this webinar, you will learn:
• Latest advancements of melt based 3D printing approaches
• Application examples for the individual technologies
• Deep dive in the MED® 3D printing technology to design dedicated drug release profiles
Presented by:
Dr. Thomas Kipping, Head of Drug Carriers
Dr. Xianghao Zuo, Deputy Director of R&D, Triastek
CAR-T Manufacturing Innovations that Work - Automating Low Volume Processes a...MilliporeSigma
Watch the presentation of this webinar here: https://bit.ly/3NDNIKe
Automated, fit-for-purpose tools are essential in CAR-T processing to support sustainable manufacturing of clinical and market-approved cell therapy products. This webinar will discuss how the ekko™ Acoustic Cell Processing System uses acoustic technology as a touchless approach to manipulate cells, enabling a modular tool across the CAR-T manufacturing workflow. Typical performance of templated ekko™ System processes for DMSO washout of leukapheresis material, low volume and high cell concentrate for electroporation preparation, and harvest of expanded T cells will be reviewed.
This webinar will also give an early glimpse at the ekko™ Select System for unmatched T cell selection.
In this webinar, you will:
• Uncover how the ekko™ System supports the broad industrialization of cell therapy, with particular focus on how to achieve low volume, high concentrate cell product for critical transduction and transfection steps
• Discover how ekko™ System for wash and concentrate processes throughout the cell therapy workflow achieve high cell recovery, viability, and effective residual removal
• Preview to ekko™ Select, our cell therapy selection platform, to achieve unmatched ease-of-use with direct processing from leukopaks reducing the need for preparation steps
Presented by:
Benjamin Ross-Johnsrud, Acoustic Technology Expert
Robert Scott, Mechanical Engineer III
How does the ICH Q5A revision impact viral safety strategies for biologics?MilliporeSigma
Watch the presentation of this webinar here: https://bit.ly/3t7X9tg
How does the ICH Q5A revision impact viral safety strategies for biologics?
Biologics continue to grow at a fast pace. Manufactured using cell lines of human or animal origin, these are at risk of viral contamination making safety strategies critical. A comprehensive risk mitigation strategy using multiple orthogonal measures is a regulatory expectation. ICH Q5A, the globally-harmonized guideline outlines the expectations. ICH Q5A is currently being revised to address recent scientific advancements including novel therapeutic modalities, new manufacturing paradigms, updates in viral clearance applications, and alternate detection technologies. We’ll discuss the expected changes and potential impact on viral safety strategies with case studies and examples.
In this webinar, you will learn about:
• The Importance of virus testing in biologics products
• Regulatory landscape, expectations for the Q5A revision
• What's new and changing
• Examples of alternate testing schedules, impact on viral clearance
Presented by:
Manjula Aysola, Senior Regulatory Consultant
Alison Armstrong, PhD, Sr. Director, Technical and Scientific Solutions
Improve Operational Efficiency by Over 30% with Product, Process, & Systems A...MilliporeSigma
Watch the presentation of this webinar here: https://bit.ly/3adaxWh
When implementing new automation systems, organizations must consider things like deployment time, user adoption, and costs.
They must also consider the cost of doing nothing – that is, what competitive advantage is lost in standing still? What time and quality is lost in repetitive, manual tasks rather than an automated, digital workflow? What operational efficiencies are lost?
In this webinar we examine how a product, process, and system agnostic automation platform can be deployed faster than traditional system specific software while bringing greater operational efficiencies (in many cases over 30% improvement).
To remain competitive in the market, biopharma manufacturers must adopt automation and digital technologies, but most plants still have island of automation consisting of independently functioning, standalone unit operations. This results in operational inefficiency, regulatory concerns, and a poor understanding of the process and product life cycle.
Taking the first, right step must include considering risks, costs, timelines, and technology alternatives. Traditional automation approaches tied to specific systems, processes, and products are, by their nature, limited; while an agnostic platform will address current biomanufacturing business challenges and ensure future readiness. With the right platform, a phased automation implementation can yield operational efficiency gains of up to 30% and improved product quality and regulatory compliance.
In this webinar, let's explore:
• Challenges of automation and digital technology adoption
• What a product, process, and system agnostic platform entails
• Applications and benefits of a process orchestration platform
• Ensuring future readiness with process orchestration
Presented by:
Braj Nandan Thakur, Global Product Manager - Automation
Insights from a Global Collaboration Accelerating Vaccine Development with an...MilliporeSigma
Watch the presentation of this webinar here: https://bit.ly/3Nbb5ug
Get insights and best practices from a multinational team establishing a platform for vaccine production. See how a long-term collaboration on a bench-scale process used to produce a Virus Like Particle (VLP) vaccine for SARS-CoV-2 was successfully converted to a robust GMP-compatible, scalable process.
The COVID-19 pandemic further emphasized the need for collaboration in the development of urgently needed vaccines and therapeutics. In this webinar, we take you behind the scenes of our collaboration with Technovax and Innovative Biotech in which a scalable VLP vaccine platform was optimized for use in a production facility in Nigeria in response to the need for local production of SARS-CoV-2 vaccines. The flexibility and robustness of the platform will enable its rapid deployment to support the West African pandemic readiness program. Initial development of the VLP process began in late 2019 and by March 2020, was already adapted for production of a SARS-CoV-2 vaccine.
In this webinar, you will learn:
• About building a priceless collaborative network with integrated solutions
• Virus-Like Particle Vaccines
• Process Development Overview and Challenges
• Pre-clinical Results and Next Steps
Presented by:
Jose M. Galarza, PhD,
President and Founder of TechnoVax
Naomi Baer,
Business development consultant, Emerging Biotech, BioProcess division
Youssef Gaabouri, Eng. ,
Associate Director, Head of Sales Middle East & Africa, BioProcess division
Risk-Based Qualification of X-Ray Sterilization for Single-Use SystemsMilliporeSigma
Watch the presentation of this webinar here: https://bit.ly/3vQf0qv
In the single-use bioprocess industry, X-ray irradiation warrants consideration as an alternate sterilization technology. Using a risk-based qualification testing strategy is important when evaluating and implementing equivalent ionizing irradiation sterilization methods.
The urgent need for life-saving therapies as a result of the global pandemic has reinforced the criticality of flexibility in pharmaceutical manufacturing, including sterilization. The single-use bioprocess industry traditionally has employed gamma irradiation sterilization. X-ray irradiation is being considered as an additional sterilization technology for business and supply continuity. We will share a risk-based qualification testing strategy including Extractables and data generated to support comparability of gamma irradiation and X-ray irradiation as equivalent ionizing irradiation sterilization methods.
In this webinar, you will learn about:
• The comparison of gamma and X-ray irradiation sterilization
• A risk-based qualification test strategy
• Data evaluation of gamma versus X-ray sterilized single-use components
Presented by:
Monica Cardona,
Global Senior Program Manager
Paul Killian, Ph.D.,
R&D Director, Analytical Technologies
Rapid Replication Competent Adenovirus (rRCA) Detection: Accelerate your Lot ...MilliporeSigma
Watch the presentation of this webinar here: https://bit.ly/3MJ4u9V
Testing for presence of replication competent adenovirus (RCA) is a key component to ensure patient safety and a requirement for all biologicals manufactured using adenoviral vectors. For many adenoviral-based products, the RCA assay is a rate-limiting assay for lot release.
Join this webinar to learn about a rapid RCA detection assay currently in development, which combines a 7-day culture assay with a highly sensitive molecular endpoint specific for RCA. The method can detect presence of as little as 1 RCA in adenoviral vector material at an approximate concentration of 5x107 - 2x108 vector particles (VP)/mL, making it a suitable method to meet regulatory requirements while accelerating your lot release timelines.
In this webinar, you will learn about:
• Regulatory framework for adenoviral vector products
• Considerations for lot release testing of adenoviral-based therapies
• Advantages of a rapid method for RCA testing on production lot material
Presented by:
Axel Fun, Ph.D.,
Principal Scientist
Alberto Santana, MBA,
Product Manager, Biologics Biosafety Testing
The High Intensity Sweeteners Neotame and Sucralose: 2 Ways to ace the Patien...MilliporeSigma
Watch the presentation of this webinar here: https://bit.ly/3vQyN7K
Bitter medicines are an important issue, especially for pediatric applications. As several APIs have bitter tasting components, high intensity sweeteners for taste optimization are of great interest. Join our webinar to discover our new sweetener toolbox enabling safe and stable formulations.
Mask bitter aftertaste for a sweeter pill to swallow! Patients’ compliance and the therapeutic benefit are supported by a pleasant taste of pharmaceutical formulations. With the high intensity sweeteners Neotame and Sucralose, you have efficient tools at hand which are superior to other sweeteners in many aspects:
• excellent sugar-like taste profile
• outstanding sweetness factors
• use effectiveness
• enhanced stability
We will present our new toolbox of two high performance sweeteners and focus on aspects of stability, safety, the application in various dosage forms, and market perception.
In this webinar, you will learn:
• How to optimize the patients' taste experience of your pharmaceuticals
• How sweeteners can be differentiated by their sensory profiles and features
• How our new product offering Neotame can be effectively used in your targeted formulations
Presented by:
Almut von der Brelie,
Senior Manager Strategic Marketing, Excipients for Solid Applications
The Developability Classification System (DCS): Enabling an Optimized Approac...MilliporeSigma
This whitepaper by Dr. Daniel Joseph Price outlines how poorly soluble drug formulations can be designed using the developability classification system (DCS).
The DCS identifies the root cause of low solubility and enables lean, cost-effective and effective formulations to be developed.
#solubility #pharmaceuticalmanufacturing #oralsoliddosage #drugdevelopment
How to Accelerate and Enhance ADC TherapiesMilliporeSigma
In this webinar, you will learn about:
The advantages of using advanced intermediates to develop ADC therapies
How to increase ADC solubility and efficiency
Fast, small-scale ADC library generation
Seamless supply chain with reduced complexity and regulatory support
The ADCore product line offers versatile intermediates that simplify the synthesis of common ADC payloads (dolastatins, maytansinoids, and PBDs) by greatly reducing the number of synthetic steps. This translates to savings in development and manufacturing costs and shorter timelines to the clinic. To address the poor solubility of many ADC payloads, ChetoSensar™ was developed to significantly increase the hydrophilicity of the drug linker, which has been shown to also substantially increase the efficacy of ADCs and broaden the therapeutic window.
Lastly, the ADC Express™ service leverages conjugation chemistry and analytical expertise to help design and quickly synthesize sets of potential ADC therapies suitable for screening to simplify candidate selection and get ADC therapies to market faster.
EU GMP Annex 1 Draft - Closed System Design Consideration with Single-Use Sys...MilliporeSigma
Biopharmaceutical manufacturing capacities have expanded dramatically which has resulted in an increased demand for single-use systems (SUS) as they have their own advantages. Although SUS are well established in the biopharmaceutical industry there is limited guidance on regulatory expectations. Please attend the webinar to learn more!
The Baculovirus Expression Vector System (BEVS) is a powerful eucaryotic vector system and Spodoptera frugiperda (Sf) cell lines are widely used as hosts for BEVS. However, the majority of Sf9 and Sf21 cell lines contain a Sf-rhabdovirus which is considered a process contaminant and must be eliminated during the process. To improve the safety profile of the BEVS production method, we offer a Sf9 rhabdovirus-negative (Sf-RVN®) cell line with companion chemically defined medium. Combined, they form the Sf-RVN® Platform which provides a performant rhabdovirus-free BEVS alternative to produce recombinant protein, VLP and AAV and enhances risk mitigation.
Addressing Downstream Challenges with Complex InjectablesMilliporeSigma
The complex injectable market is gaining traction in the injectable therapies, however manufacturing of it is critical. In this webinar, lets brainstorm on the downstream criticalities of these molecules and how to handle the same.
Earliest Galaxies in the JADES Origins Field: Luminosity Function and Cosmic ...Sérgio Sacani
We characterize the earliest galaxy population in the JADES Origins Field (JOF), the deepest
imaging field observed with JWST. We make use of the ancillary Hubble optical images (5 filters
spanning 0.4−0.9µm) and novel JWST images with 14 filters spanning 0.8−5µm, including 7 mediumband filters, and reaching total exposure times of up to 46 hours per filter. We combine all our data
at > 2.3µm to construct an ultradeep image, reaching as deep as ≈ 31.4 AB mag in the stack and
30.3-31.0 AB mag (5σ, r = 0.1” circular aperture) in individual filters. We measure photometric
redshifts and use robust selection criteria to identify a sample of eight galaxy candidates at redshifts
z = 11.5 − 15. These objects show compact half-light radii of R1/2 ∼ 50 − 200pc, stellar masses of
M⋆ ∼ 107−108M⊙, and star-formation rates of SFR ∼ 0.1−1 M⊙ yr−1
. Our search finds no candidates
at 15 < z < 20, placing upper limits at these redshifts. We develop a forward modeling approach to
infer the properties of the evolving luminosity function without binning in redshift or luminosity that
marginalizes over the photometric redshift uncertainty of our candidate galaxies and incorporates the
impact of non-detections. We find a z = 12 luminosity function in good agreement with prior results,
and that the luminosity function normalization and UV luminosity density decline by a factor of ∼ 2.5
from z = 12 to z = 14. We discuss the possible implications of our results in the context of theoretical
models for evolution of the dark matter halo mass function.
(May 29th, 2024) Advancements in Intravital Microscopy- Insights for Preclini...Scintica Instrumentation
Intravital microscopy (IVM) is a powerful tool utilized to study cellular behavior over time and space in vivo. Much of our understanding of cell biology has been accomplished using various in vitro and ex vivo methods; however, these studies do not necessarily reflect the natural dynamics of biological processes. Unlike traditional cell culture or fixed tissue imaging, IVM allows for the ultra-fast high-resolution imaging of cellular processes over time and space and were studied in its natural environment. Real-time visualization of biological processes in the context of an intact organism helps maintain physiological relevance and provide insights into the progression of disease, response to treatments or developmental processes.
In this webinar we give an overview of advanced applications of the IVM system in preclinical research. IVIM technology is a provider of all-in-one intravital microscopy systems and solutions optimized for in vivo imaging of live animal models at sub-micron resolution. The system’s unique features and user-friendly software enables researchers to probe fast dynamic biological processes such as immune cell tracking, cell-cell interaction as well as vascularization and tumor metastasis with exceptional detail. This webinar will also give an overview of IVM being utilized in drug development, offering a view into the intricate interaction between drugs/nanoparticles and tissues in vivo and allows for the evaluation of therapeutic intervention in a variety of tissues and organs. This interdisciplinary collaboration continues to drive the advancements of novel therapeutic strategies.
Cancer cell metabolism: special Reference to Lactate PathwayAADYARAJPANDEY1
Normal Cell Metabolism:
Cellular respiration describes the series of steps that cells use to break down sugar and other chemicals to get the energy we need to function.
Energy is stored in the bonds of glucose and when glucose is broken down, much of that energy is released.
Cell utilize energy in the form of ATP.
The first step of respiration is called glycolysis. In a series of steps, glycolysis breaks glucose into two smaller molecules - a chemical called pyruvate. A small amount of ATP is formed during this process.
Most healthy cells continue the breakdown in a second process, called the Kreb's cycle. The Kreb's cycle allows cells to “burn” the pyruvates made in glycolysis to get more ATP.
The last step in the breakdown of glucose is called oxidative phosphorylation (Ox-Phos).
It takes place in specialized cell structures called mitochondria. This process produces a large amount of ATP. Importantly, cells need oxygen to complete oxidative phosphorylation.
If a cell completes only glycolysis, only 2 molecules of ATP are made per glucose. However, if the cell completes the entire respiration process (glycolysis - Kreb's - oxidative phosphorylation), about 36 molecules of ATP are created, giving it much more energy to use.
IN CANCER CELL:
Unlike healthy cells that "burn" the entire molecule of sugar to capture a large amount of energy as ATP, cancer cells are wasteful.
Cancer cells only partially break down sugar molecules. They overuse the first step of respiration, glycolysis. They frequently do not complete the second step, oxidative phosphorylation.
This results in only 2 molecules of ATP per each glucose molecule instead of the 36 or so ATPs healthy cells gain. As a result, cancer cells need to use a lot more sugar molecules to get enough energy to survive.
Unlike healthy cells that "burn" the entire molecule of sugar to capture a large amount of energy as ATP, cancer cells are wasteful.
Cancer cells only partially break down sugar molecules. They overuse the first step of respiration, glycolysis. They frequently do not complete the second step, oxidative phosphorylation.
This results in only 2 molecules of ATP per each glucose molecule instead of the 36 or so ATPs healthy cells gain. As a result, cancer cells need to use a lot more sugar molecules to get enough energy to survive.
introduction to WARBERG PHENOMENA:
WARBURG EFFECT Usually, cancer cells are highly glycolytic (glucose addiction) and take up more glucose than do normal cells from outside.
Otto Heinrich Warburg (; 8 October 1883 – 1 August 1970) In 1931 was awarded the Nobel Prize in Physiology for his "discovery of the nature and mode of action of the respiratory enzyme.
WARNBURG EFFECT : cancer cells under aerobic (well-oxygenated) conditions to metabolize glucose to lactate (aerobic glycolysis) is known as the Warburg effect. Warburg made the observation that tumor slices consume glucose and secrete lactate at a higher rate than normal tissues.
A brief information about the SCOP protein database used in bioinformatics.
The Structural Classification of Proteins (SCOP) database is a comprehensive and authoritative resource for the structural and evolutionary relationships of proteins. It provides a detailed and curated classification of protein structures, grouping them into families, superfamilies, and folds based on their structural and sequence similarities.
Multi-source connectivity as the driver of solar wind variability in the heli...Sérgio Sacani
The ambient solar wind that flls the heliosphere originates from multiple
sources in the solar corona and is highly structured. It is often described
as high-speed, relatively homogeneous, plasma streams from coronal
holes and slow-speed, highly variable, streams whose source regions are
under debate. A key goal of ESA/NASA’s Solar Orbiter mission is to identify
solar wind sources and understand what drives the complexity seen in the
heliosphere. By combining magnetic feld modelling and spectroscopic
techniques with high-resolution observations and measurements, we show
that the solar wind variability detected in situ by Solar Orbiter in March
2022 is driven by spatio-temporal changes in the magnetic connectivity to
multiple sources in the solar atmosphere. The magnetic feld footpoints
connected to the spacecraft moved from the boundaries of a coronal hole
to one active region (12961) and then across to another region (12957). This
is refected in the in situ measurements, which show the transition from fast
to highly Alfvénic then to slow solar wind that is disrupted by the arrival of
a coronal mass ejection. Our results describe solar wind variability at 0.5 au
but are applicable to near-Earth observatories.
Slide 1: Title Slide
Extrachromosomal Inheritance
Slide 2: Introduction to Extrachromosomal Inheritance
Definition: Extrachromosomal inheritance refers to the transmission of genetic material that is not found within the nucleus.
Key Components: Involves genes located in mitochondria, chloroplasts, and plasmids.
Slide 3: Mitochondrial Inheritance
Mitochondria: Organelles responsible for energy production.
Mitochondrial DNA (mtDNA): Circular DNA molecule found in mitochondria.
Inheritance Pattern: Maternally inherited, meaning it is passed from mothers to all their offspring.
Diseases: Examples include Leber’s hereditary optic neuropathy (LHON) and mitochondrial myopathy.
Slide 4: Chloroplast Inheritance
Chloroplasts: Organelles responsible for photosynthesis in plants.
Chloroplast DNA (cpDNA): Circular DNA molecule found in chloroplasts.
Inheritance Pattern: Often maternally inherited in most plants, but can vary in some species.
Examples: Variegation in plants, where leaf color patterns are determined by chloroplast DNA.
Slide 5: Plasmid Inheritance
Plasmids: Small, circular DNA molecules found in bacteria and some eukaryotes.
Features: Can carry antibiotic resistance genes and can be transferred between cells through processes like conjugation.
Significance: Important in biotechnology for gene cloning and genetic engineering.
Slide 6: Mechanisms of Extrachromosomal Inheritance
Non-Mendelian Patterns: Do not follow Mendel’s laws of inheritance.
Cytoplasmic Segregation: During cell division, organelles like mitochondria and chloroplasts are randomly distributed to daughter cells.
Heteroplasmy: Presence of more than one type of organellar genome within a cell, leading to variation in expression.
Slide 7: Examples of Extrachromosomal Inheritance
Four O’clock Plant (Mirabilis jalapa): Shows variegated leaves due to different cpDNA in leaf cells.
Petite Mutants in Yeast: Result from mutations in mitochondrial DNA affecting respiration.
Slide 8: Importance of Extrachromosomal Inheritance
Evolution: Provides insight into the evolution of eukaryotic cells.
Medicine: Understanding mitochondrial inheritance helps in diagnosing and treating mitochondrial diseases.
Agriculture: Chloroplast inheritance can be used in plant breeding and genetic modification.
Slide 9: Recent Research and Advances
Gene Editing: Techniques like CRISPR-Cas9 are being used to edit mitochondrial and chloroplast DNA.
Therapies: Development of mitochondrial replacement therapy (MRT) for preventing mitochondrial diseases.
Slide 10: Conclusion
Summary: Extrachromosomal inheritance involves the transmission of genetic material outside the nucleus and plays a crucial role in genetics, medicine, and biotechnology.
Future Directions: Continued research and technological advancements hold promise for new treatments and applications.
Slide 11: Questions and Discussion
Invite Audience: Open the floor for any questions or further discussion on the topic.
This pdf is about the Schizophrenia.
For more details visit on YouTube; @SELF-EXPLANATORY;
https://www.youtube.com/channel/UCAiarMZDNhe1A3Rnpr_WkzA/videos
Thanks...!
The increased availability of biomedical data, particularly in the public domain, offers the opportunity to better understand human health and to develop effective therapeutics for a wide range of unmet medical needs. However, data scientists remain stymied by the fact that data remain hard to find and to productively reuse because data and their metadata i) are wholly inaccessible, ii) are in non-standard or incompatible representations, iii) do not conform to community standards, and iv) have unclear or highly restricted terms and conditions that preclude legitimate reuse. These limitations require a rethink on data can be made machine and AI-ready - the key motivation behind the FAIR Guiding Principles. Concurrently, while recent efforts have explored the use of deep learning to fuse disparate data into predictive models for a wide range of biomedical applications, these models often fail even when the correct answer is already known, and fail to explain individual predictions in terms that data scientists can appreciate. These limitations suggest that new methods to produce practical artificial intelligence are still needed.
In this talk, I will discuss our work in (1) building an integrative knowledge infrastructure to prepare FAIR and "AI-ready" data and services along with (2) neurosymbolic AI methods to improve the quality of predictions and to generate plausible explanations. Attention is given to standards, platforms, and methods to wrangle knowledge into simple, but effective semantic and latent representations, and to make these available into standards-compliant and discoverable interfaces that can be used in model building, validation, and explanation. Our work, and those of others in the field, creates a baseline for building trustworthy and easy to deploy AI models in biomedicine.
Bio
Dr. Michel Dumontier is the Distinguished Professor of Data Science at Maastricht University, founder and executive director of the Institute of Data Science, and co-founder of the FAIR (Findable, Accessible, Interoperable and Reusable) data principles. His research explores socio-technological approaches for responsible discovery science, which includes collaborative multi-modal knowledge graphs, privacy-preserving distributed data mining, and AI methods for drug discovery and personalized medicine. His work is supported through the Dutch National Research Agenda, the Netherlands Organisation for Scientific Research, Horizon Europe, the European Open Science Cloud, the US National Institutes of Health, and a Marie-Curie Innovative Training Network. He is the editor-in-chief for the journal Data Science and is internationally recognized for his contributions in bioinformatics, biomedical informatics, and semantic technologies including ontologies and linked data.
Observation of Io’s Resurfacing via Plume Deposition Using Ground-based Adapt...Sérgio Sacani
Since volcanic activity was first discovered on Io from Voyager images in 1979, changes
on Io’s surface have been monitored from both spacecraft and ground-based telescopes.
Here, we present the highest spatial resolution images of Io ever obtained from a groundbased telescope. These images, acquired by the SHARK-VIS instrument on the Large
Binocular Telescope, show evidence of a major resurfacing event on Io’s trailing hemisphere. When compared to the most recent spacecraft images, the SHARK-VIS images
show that a plume deposit from a powerful eruption at Pillan Patera has covered part
of the long-lived Pele plume deposit. Although this type of resurfacing event may be common on Io, few have been detected due to the rarity of spacecraft visits and the previously low spatial resolution available from Earth-based telescopes. The SHARK-VIS instrument ushers in a new era of high resolution imaging of Io’s surface using adaptive
optics at visible wavelengths.
The Viscosity Reduction Platform: Viscosity-Reducing Excipients for Protein Formulation
1. Introduction
Protein viscosity is one of the major obstacles in preparing highly
concentrated protein formulations suitable for subcutaneous
(subQ) injection. Highly viscous protein solutions would require
a significant force to be applied to the syringe for injection. As
a result, the patient could experience a considerable amount of
pain. In many cases, injectability would not be possible.1,2
The Viscosity Reduction
Platform: Viscosity-
reducing excipients
for protein formulation
Authors: Stefan Braun, Niels Banik, Jennifer J. Widera,
Jan Gerit Brandenburg and Tobias Rosenkranz
0
200
150
100
50
0 50 100
Viscosity
[mPa·s]
Concentration [mg/mL]
150 200 250 300
Limit of
injectability
Interactions Crowding
mAbA
mAbB
mAbC
mAbD
mAbE
mAbF
mAbG
mAbH
mAbL
mAbJ
mAbK
Concentration dependency of
viscosity for market formulations
Figure 1.
Dependency of viscosity on antibody concentration and likely underlying causes.
Red lines show injectability limit.
White Paper
When characterizing protein viscosity
behavior, one can differentiate two different
concentration regimes as shown in Figure 1.
At very low concentrations below about
75 mg/mL, proteins are rarely viscous. When
increasing the concentration to between
100 and 200 mg/mL, some proteins exhibit
elevated viscosity exceeding the limit of
injectability, which is typically between
20 and 25 mPa·s. At this concentration
regime, several proteins exhibit an affinity
for self-interaction, i.e. forming transient
clusters that give rise to elevated viscosity.
At concentrations above 200 mg/mL, the
nearest neighbor distance between the
protein molecules shrinks so that without
a specific affinity for self-interactions, said
protein-protein interactions take place. While
viscosity-reducing excipients can affect
proteins exhibiting either of these interaction
patterns, they are likely to be more efficient
at protein concentration regimes below
200 mg/mL.3,4
MilliporeSigma is the U.S. and
Canada Life Sciencebusiness of
Merck KGaA, Darmstadt, Germany.
2. 2
These intermolecular interactions between proteins
have the same molecular origin as the intramolecular
interactions that structurally stabilize the proteins.
This means viscosity-reducing excipients that affect
protein-protein interactions can potentially also
destabilize proteins. As such, it is essential to balance
an excipient’s viscosity-reducing ability against its
potential to destabilize a protein. For some excipients,
a concentration-dependent effect on protein stability
is well-documented. At lower concentrations, the
excipients act as stabilizers, but this behavior changes
as concentration increases, often with an adverse
effect on protein stability. Excipient concentration
is thus a critical factor in managing protein stability.
These two aspects can be better balanced by using an
excipient combination of an amino acid and an anionic
excipient. When used in combination, excipients are
more efficient in reducing viscosity and may even do
so in an over-additive manner. Consequently, lower
concentrations of the individual excipients can be used,
which is more favorable for protein stability.
This white paper evaluates the viscosity-reducing
capacities of excipients and excipient combinations. It
shows the over-additive effect of using two excipients
together and addresses how excipients’ viscosity-
reducing ability depends on pH. The results show
the effect of protein viscosity on injection force and
highlight the platform’s ability to balance viscosity
reduction with protein stability. The case studies
presented demonstrate that using a combination of
two excipients at lower concentrations instead of a
single excipient at a higher concentration enables
balancing protein viscosity and protein stability in
a favorable way.
Table 1: Excipients and abbreviations
Excipient Abbrev.
L-Ornithine monohydrochloride Orn
L-Phenylalanine Phe
Thiamine phosphoric acid ester chloride dihydrate TMP
Benzenesulfonic acid BSAcid
Pyridoxine hydrochloride Pyr
Results & Discussion
Table 1 summarizes the excipients that are part of
the Viscosity Reduction Platform. For clarity reasons,
abbreviations mentioned in Table 1 are used in the
fol
lowing. The benchmark excipient is referred to as BM.
Single excipients often reduce viscosity
but may impact protein stability
A single excipient is often used to reduce the viscosity
of a protein formulation. Figure 2 shows two model
proteins, infliximab and evolocumab, where each
component of the Viscosity Reduction Platform has
been used individually. Infliximab has a viscosity of
about 40 mPa·s at a concentration of 120 mg/mL in
its concentrated marketed formulation (see Figure
2A). Adding 75 mM of the single excipients reduces
the viscosity by anywhere from 10 to 80%. A similar
viscosity reduction is observed when doubling the
excipient concentration to 150 mM. Comparing the
performance of an excipient at 75 and 150 mM shows
that the greatest difference in viscosity reduction
between the two concentrations is seen with excipients
that are not particularly effective. Excipients able
to halve the viscosity of infliximab do not show a
proportionally strong viscosity-reducing effect when
their concentration is increased. Used individually, BM
and Orn do not reduce infliximab viscosity effectively.
However, we will show that these two excipients
can indeed be valuable when used in excipient
combinations.
0
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60
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Viscosity
[mPa·s]
Infliximab
Control
75 mM
150 mM
A
0
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60
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Viscosity
[mPa·s]
Evolocumab
Control
75 mM
150 mM
B
Figure 2.
Influence of increased excipient concentrations on protein formulation viscosity.
3. 3
Similarly, Figure 2B shows that many excipients lead
to an improved viscosity reduction for 170 mg/mL
evolocumab when used at higher concentrations.
By contrast, Phe actually increases protein viscosity
when its concentration is increased. Overall, for both
model antibodies, it was observed that doubling the
concentration of an excipient does not typically lead
to improved viscosity reduction.
Balancing viscosity reduction and protein stability
is crucial to successfully develop a stable, highly
concentrated protein formulation. A forced degradation
study was thus conducted to evaluate the effect of
elevated excipient concentrations (125–150 mM) on
protein stability. Figure 3 summarizes the monomer
content of infliximab and evolocumab formulations
after 28 days at 40 °C and 75% relative humidity.
Infliximab was formulated at a concentration of
120 mg/mL, while evolocumab was formulated at a
concentration of 170 mg/mL. The amino acids do not
show an adverse effect on protein stability, with the
exception of Phe, which is the most effective viscosity-
reducing amino acid for infliximab. Phe’s observed
destabilizing effect highlights the importance of
bal
ancing protein stability and protein viscosity.
0
50
75
25
100
125
C
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M
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Infliximab Evolocumab
Residual
Monomer
Content
[%]
Figure 3.
Effect of single excipients at concentrations between 125–150 mM
on monomer content of infliximab and evolocumab stored at 40 °C/
75% rH for 28 days.
The three anionic excipients show a clear destabilizing
effect on both proteins, as can be seen in Figure 3.
With TMP, a substantial loss of monomer content is
seen, likely due to the known instability of the vitamin
derivate itself at high temperatures. In summary,
highly efficient viscosity-reducing excipients used
at concentrations between 125 mM and 150 mM
can desta
bilize a protein. In contrast, amino acids
typi
cally allow protein stability to be maintained.
To conclude, while increasing the excipient concen
tration may allow for improved viscosity reduction,
some excipients can destabilize proteins when used
at high concentrations. Furthermore, even these
increased excipient concentrations may not be able
to lower viscosity sufficiently to reach the targeted
formulation viscosity.
Effect of protein formulation pH on excipient
performance
As demonstrated, excipients’ viscosity-reducing ability
can differ depending on the protein they are used
for. As a next step, it is important to consider the
formulation conditions. Figure 4 shows the viscosity
of 170 mg/mL evolocumab formulated at pH 5 (acetate
buffer) and pH 7.2 (phosphate buffer). The materials
used to prepare the base buffer are listed in Table 2.
Table 2: Materials used for base buffer preparation
Buffer Buffer Components
Acetate
buffer
Acetic acid (glacial) 100% EMPROVE®
EXPERT Ph Eur,
BP,JP,USP
Sodium hydroxide solution 32%
EMPROVE®
EXPERT
Phosphate
buffer
Sodium dihydrogen phosphate monohydrate
EMPROVE®
EXPERT BP,USP
di-Sodium hydrogen phosphate heptahydrate
EMPROVE®
EXPERT DAC,USP
Optional addition: Sodium chloride EMPROVE®
EXPERT
Ph Eur,BP,ChP,JP,USP
0
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80
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pH 7.2
pH 5.0
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Viscosity
[mPa·s]
pH-dependent excipient performance
Figure 4.
pH dependency of evolocumab formulation at 170 mg/mL:
Comparison using pH 5 acetate and pH 7.2 phosphate buffers.
4. 4
Table 3: Physical properties of excipient molecules at pH 5 and pH 7
Charge
[atomic units]
Dipole moment
[Debye]
Solvent-accessible
surface area (SASA) [Å2
]
MolLogP
Excipient pH 5.0 pH 7.2 pH 5.0 pH 7.2 pH 5.0 pH 7.2 pH 5.0 pH 7.2
L-Ornithine hydrochloride 1.0 1.0 25.5 25.5 304 304 –5.3 –5.3
L-Phenylalanine 0.0 0.0 11.3 11.3 275 275 –1.4 –1.4
Benzenesulfonic acid –1.0 –1.0 13.7 13.7 231 231 0.6 0.6
Thiamine phosphoric acid ester chloride –0.2 –1.7 29.6 26.2 442 469 –3.7 –4.9
Benchmark 1.0 1.0 9.0 9.1 290 289 –5.5 –5.5
Pyridoxine 0.8 0.0 3.3 3.7 276 274 –0.4 0.1
Formulated in the respective base buffers, the viscosity
is much higher than 20 mPa·s. At pH 5.0, it is 59 mPa·s,
and at pH 7.2, it is 72 mPa·s. Adding sodium chloride
has a stronger effect at pH 7.2 than at pH 5, potentially
due to the lower number of charges present on the
protein at pH 7.2, which is closer to the protein’s
isoelectric point of about 7.6. The Viscosity Reduction
Platform excipients (see Table 1) show differing trends.
The performance of Phe is stable with respect to the
pH condition. The excipients BM, Orn, BSAcid, Pyr, and
TMP exhibit changes in performance at different pH
levels. Computational chemistry techniques were used
to calculate a selection of relevant excipient properties
across a pH range of 4 to 8. These parameters were
used to determine whether this difference in viscosity
reduction could be explained by changes in excipient
or protein properties. The underlying molecular pKa
values significantly impact these properties and were
confirmed experimentally by titration studies.
A summary is given in Table 3.
Only in the case of TMP a change in protonation
state was found when the pH was reduced to 5.
Accordingly, changes were observed in dipole
moment, accessible surface area, and the water-
octanol partition coefficient indicating the molecule’s
hydrophobicity. TMP was nevertheless a highly
efficient viscosity-reducing excipient for evolocumab
under both formulation conditions. As there is no pH-
dependent change for the other excipient molecules,
the difference in viscosity-reducing performance with
evolocumab likely has a protein origin. Evolocumab’s
hydrophobicity is pH-independent, leading to an
increased charge on the protein at a lower pH, which
affects protein-excipient interactions. This case study
suggests that different excipients may be required
to formulate a protein under different conditions.
An excipient toolbox would thus allow formulation
scientists to find the right excipients for the desired
formulation conditions.
Using excipient combinations to reduce
protein viscosity
As individual excipients may not be powerful enough
to reduce the viscosity of a highly concentrated protein
formulation on their own, the Viscosity Reduction
Platform is based on the use of excipient combinations.
An amino acid – i.e. BM, Orn or Phe – is combined
with an anionic excipient. Being able to vary excipient
combinations in this way gives formulation scientists
a high degree of flexibility when it comes to balancing
viscosity reduction against protein stability and other
considerations like route of administration, which
may determine the pH of the formulation that is to
be developed.
5. 5
Figure 5A shows the formulation viscosity of infliximab
at a concentration of 120 mg/mL with a variety of
excipient combinations. The grey control bar is the
unmodified marketed formulation concentrated to the
given protein concentration. The resulting viscosity
of about 40 mPa·s is too high for subcutaneous
administration. The purple bar represents the
benchmark excipient, which by itself is only able
to slightly reduce the viscosity. In several cases,
combining Orn, BM or Phe with an anionic excipient
leads to a more substantial reduction in viscosity
– including below the injectability limit, most
importantly. With each amino acid, there are multiple
combinations that would allow for injectability of
infliximab. Orn is particularly effective with Pyr. The
benchmark excipient can be combined with TMP. Phe
is best combined with BSAcid or TMP. In summary,
different excipient combinations are efficient for
infliximab. However, not all excipients may be suitable
for every route of administration due to potential
tissue-specific reactions, which is why using an
excipient portfolio is beneficial.
Figure 5B shows the same approach using evolocumab
as a model protein. Here, 150 mM of sodium chloride
was included as a control to monitor ionic effects. In
contrast to infliximab, evolocumab is marketed in a
low-salt formulation. Evolocumab’s viscosity can be
managed well with the benchmark excipient. However,
there are conditions where the benchmark excipient
is not desirable because of the route of administration
or a local reaction to the excipient. The Viscosity
Reduction Platform presented here provides a range
of alternatives.
0
20
60
Viscosity
[mPa·s]
120 mg/mL Infliximab
40
Injectability
Limit
BM
Orn
Pyr
Phe
BSAcid TMP
Control
A
Injectability
Limit
170 mg/mL Evolocumab
Control
TMP
0
20
60
40
Viscosity
[mPa·s]
BM
NaCl
BSAcid
Orn
Phe Pyr
B
Figure 5.
Combinations of Viscosity Reduction Platform excipients compared to experiments without a viscosity-reducing excipient (grey bar), with sodium
chloride as control (green bar) and the industry standard as benchmark (purple bar). The color codes of the split bars indicate the excipient
combinations used. A) Model antibody infliximab. B) Model antibody evolocumab.
To further illustrate the potential benefits of using
excipient combinations, their performance was
assessed with respect to over-additive effects. Figure
6 shows the measured viscosity for each excipient
combination versus the expected viscosity for that
combination based on measurements of formulations
with the single excipient alone. Data points below
the identity line indicate an over-additive viscosity-
reducing effect, which is seen with several excipient
combinations. Others, however, result in a decrease
in viscosity yet do not display an over-additive effect.
This behavior likely depends on the protein in question
and the formulation conditions.
0
60
40
20
20 40 60 80 100
0
80
100
Measured
Viscosity
[mPa·s]
Calculated Viscosity [mPa·s]
Over-additive
Orn/BSAcid
Phe/BSAcid
Infliximab
Evolocumab
BM/Pyr
BM/BSAcid
Orn/BSAcid
Figure 6.
Over-additive effect of excipient combinations on viscosity reduction.
6. 6
In summary, the data with these two model antibodies
shows that using excipient combinations can reduce
viscosity more effectively than the leading industry
standard.
Combined excipients are also more efficient than
highly concentrated single excipients and can even
perform synergistically. Moreover, an excipient
portfolio gives formulation scientists greater flexibility.
Depending on the nature of the antibody, the desired
pH, or the route of administration, having a variety
of options at hand can be beneficial when developing
the final formulation. The most suitable choice of
excipients will depend on the type of protein and the
formulation conditions.
Impact of reduced protein viscosity on
syringeability
To highlight the impact of viscosity and the Viscosity
Reduction Platform on syringeability, the following case
study investigates two relevant factors: aspiration
time and extraction force. First, the aspiration time
of infliximab and evolocumab was tested at high
concentrations (120 mg/mL and 170 mg/mL) with
and without the most effective viscosity-reducing
excipients (Figure 7A). Aspirating infliximab into a 1 mL
syringe through a 27-gauge needle takes 75 s. With
Orn/TMP this time can be reduced by 19%, and with
Phe/TMP by 44%. For evolocumab, it takes 116 s to
aspirate a highly concentrated solution into the same
syringe. With Orn/Pyr this time can be reduced to
46 s, and with BM/TMP to 37 s.
Figure 7B shows the syringe extraction force required
for different formulations of infliximab and evolocumab
using a 1 mL syringe through a 27-gauge needle (BD
Plastipak™ 1 mL syringe, 27G, 13 mm needle). The
syringe extraction force is very sensitive to the type
of syringe used, its dimension, the needle length,
and the inner needle diameter. In the present study
a flow rate of 0.2 mL/s is used to showcase the
impact of the Viscosity Reduction Platform on the
injection force. Flow rates of 0.15 mL/s and 0.45 mL/s
are described in literature.7
Evolocumab is supplied
by the manufacturer in a pen to self-inject using a
flow rate of 0.2 mL/s. Therefore this flow rate was
chosen as an example. An extraction force of about
20 N was observed for 120 mg/mL infliximab in its
marketed formulation. Viscosity-reducing excipients
can reduce this to about 15 N. For 170 mg/mL
evolocumab, the difference is even more pronounced.
In the standard buffer, an extraction force of 30 N
was measured. Both excipient combinations are able
to reduce the extraction force by about 50%. These
examples highlight the practical impact that reduced
formulation viscosity has on the syringeability of highly
concentrated protein solutions.
Control
0
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30
90
120
O
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Time
[s]
Aspiration time
B
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Infliximab Evolocumab
A
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Force
[N]
Extraction force
B
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Control
Control
Infliximab Evolocumab
B
Figure 7.
A) The aspiration time of infliximab and evolocumab in their reference
buffers versus formulated with the best-performing viscosity-reducing
excipient combinations, and B) the extraction force of the two
molecules in the same formulation.
7. 7
0
25
100
125
Monomer
Content
[%]
Monomer content of Infliximab
stabilized by excipient combinations
75
50
Control
Orn
Phe
BSAcid
Pyr
TMP
BM
A
Monomer
Content
[%]
Monomer content of Evolocumab
stabilized by using combinations
Control
Orn
Phe
BSAcid
Pyr
TMP
BM
0
25
100
125
75
50
B
Figure 8.
Monomer content of A) infliximab and B) evolocumab formulations after a forced degradation study of 28 days at 40 °C/75% rH. Solid bars
represent data with only one excipient, split bars represent excipient combinations, where the color code indicates which excipients were used.
Addressing protein stability with the
Viscosity Reduction Platform
As previously discussed, the balance between protein
viscosity and protein stability is rather delicate.
Focusing on protein stability, a forced degradation
study was performed using combinations of excipients
with varying concentrations of the individual compo
nents. As shown in Figure 8, excipient combinations
can overcome the adverse effect of using an anionic
excipient alone. The formulations used were not
optimized further after addition of the viscosity-
reducing excipients. Instead, the stability of the
two model proteins was investigated over a longer
period at 2–8 °C and 25 °C/60% relative humidity.
Figure 9A shows for all selected excipient combinations
that infliximab and evolocumab were able to retain a
high monomer content after 24 weeks at 2–8 °C. This
Start 8 weeks 16 weeks 24 weeks
C
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Monomer
Content
[%]
Stability when stored at 25 °C/60% rH
Infliximab Evolocumab
B
Figure 9.
Long-term stability of selected formulations with excipient combinations that successfully reduced viscosity. A) Stability at 2–8 °C and B) stability
at 25 °C/60% rH.
high stability was achieved without further optimization
of the formulation and could thus be potentially
improved even more if the antibody were to undergo
thorough formulation development. It is particularly
noteworthy that the combination of Phe and TMP is
able to maintain a high monomer content at 2–8 °C.
At 25 °C, formulations containing TMP showed a strong
destabilizing effect up to a total loss of monomer. This
further supports the hypothesis that the decrease in
protein stability is due to the decomposition of the
excipient molecule. When stored under accelerated
conditions, i.e. 25 °C/60% rH, a high monomer content
(even above 95% in some cases) was observed for
selected excipient combinations. Overall, it was shown
that using viscosity-reducing excipients in combination
with each other can maintain formulation stability under
relevant storage conditions.
C
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Stability when stored at 2–8°C
P
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0
50
100
150
Monomer
Content
[%]
Infliximab Evolocumab
Start 8 weeks 16 weeks 24 weeks
A