Biopharmaceutical include vast range of proteins which can be modified at a molecular level and used in artificial preparation of vaccines, nutraceutical products and various products on a scale. Biopharmaceuticals are usually derivatives of natural human proteins, which make them ideal for targeted cellular therapy. Unlike small molecule drugs, which penetrate cell membranes including healthy cells, Biopharmaceuticals act through external cellular binding to induce the desired cellular response.
Biopharmaceutical include vast range of proteins which can be modified at a molecular level and used in artificial vaccines, nutraceutical products and various product on a large scale.
Designing of drug delivery system for biotechnology products considering stab...Smaranika Rahman
"Where there is life, there is DNA, where there is DNA, there is biotechnology." Biotechnology, as the word suggests, is combination of biology and technology. So the importance of biotechnology and biotechnology products in our life is increasing day by day. That's why we have to produce biotechnology products in a safer manner and also maintain that through it's shelf-life.We have also research on improving methods of improving it's stability. In this topic, I also tried to discuss bioinformatic-driven strategies that are used to predict structural changes that can be applied to wild type proteins in order to produce more stable variants. The most commonly employed techniques PEGylation, stochastic approaches, empirical or systematic rational design strategies.
Agarikon.1 and Agarikon Plus Affect Cell Cycle and Induce Apoptosis in Human ...Neven Jakopovic
An investigation into the antiproliferative and pro-apoptotic mechanisms of medicinal mushroom extract blends (Myko San company) on human lung and colon carcinoma cell lines. This is a continuation of a study conducted in 2013 (Durgo, Jakopovich)
It was established that these products possess antiproliferative, mainly cytostatic activity; induce cell cycle preturbations by delaying the progress through the G1 and S phase pointing to disturbances before or during DNA replication; exhibit mild early and late apoptosis; and cause non-specific cytotoxic effect on cancer cells.
The results were presented by Boris Jakopovich, at the 7th International Medicinal Mushroom Conference in Beijing, China, in 2013.
The main focus of discovery biology for the past few years has been in the areas of diabetes, cardiovascular, cancer, infl ammation and anti-infective. One of the key out of box approach for combating diabetes has been the regulation of key protein AMP kinase. For cancer therapy, several of the signal transduction kinases like ERK kinase, MAP kinase, Aurora kinase etc have been used as key protein targets. Kinases are enzymes that regulate downstream pathways by post translational modifi cation (phosphorylation) of target proteins. Thus the detection of phosphorylation state of a protein is very important in these studies. Classically kinase assays were and are performed using radiolabeled P32 or P33 antibodies that have serious disadvantages. The regulation of proteins at the gene and mRNA level can be very well studied by genomics technology but the post translational modifi cation of proteins can be studied exclusively by proteomic strategies. In view of the tremendous importance of kinases to discovery research, a non-radioactive, non-antibody, high throughput and safe assay for detection of in vivo phosphorylation of proteins -the proteomic way is proposed.
Biopharmaceutical include vast range of proteins which can be modified at a molecular level and used in artificial vaccines, nutraceutical products and various product on a large scale.
Designing of drug delivery system for biotechnology products considering stab...Smaranika Rahman
"Where there is life, there is DNA, where there is DNA, there is biotechnology." Biotechnology, as the word suggests, is combination of biology and technology. So the importance of biotechnology and biotechnology products in our life is increasing day by day. That's why we have to produce biotechnology products in a safer manner and also maintain that through it's shelf-life.We have also research on improving methods of improving it's stability. In this topic, I also tried to discuss bioinformatic-driven strategies that are used to predict structural changes that can be applied to wild type proteins in order to produce more stable variants. The most commonly employed techniques PEGylation, stochastic approaches, empirical or systematic rational design strategies.
Agarikon.1 and Agarikon Plus Affect Cell Cycle and Induce Apoptosis in Human ...Neven Jakopovic
An investigation into the antiproliferative and pro-apoptotic mechanisms of medicinal mushroom extract blends (Myko San company) on human lung and colon carcinoma cell lines. This is a continuation of a study conducted in 2013 (Durgo, Jakopovich)
It was established that these products possess antiproliferative, mainly cytostatic activity; induce cell cycle preturbations by delaying the progress through the G1 and S phase pointing to disturbances before or during DNA replication; exhibit mild early and late apoptosis; and cause non-specific cytotoxic effect on cancer cells.
The results were presented by Boris Jakopovich, at the 7th International Medicinal Mushroom Conference in Beijing, China, in 2013.
The main focus of discovery biology for the past few years has been in the areas of diabetes, cardiovascular, cancer, infl ammation and anti-infective. One of the key out of box approach for combating diabetes has been the regulation of key protein AMP kinase. For cancer therapy, several of the signal transduction kinases like ERK kinase, MAP kinase, Aurora kinase etc have been used as key protein targets. Kinases are enzymes that regulate downstream pathways by post translational modifi cation (phosphorylation) of target proteins. Thus the detection of phosphorylation state of a protein is very important in these studies. Classically kinase assays were and are performed using radiolabeled P32 or P33 antibodies that have serious disadvantages. The regulation of proteins at the gene and mRNA level can be very well studied by genomics technology but the post translational modifi cation of proteins can be studied exclusively by proteomic strategies. In view of the tremendous importance of kinases to discovery research, a non-radioactive, non-antibody, high throughput and safe assay for detection of in vivo phosphorylation of proteins -the proteomic way is proposed.
Overview Radboudumc Center for Proteomics, Glycomics and Metabolomics april 2015Alain van Gool
An overview of the proteomics, glycomics and metabolomics expertise and capabilities within the Translational Metabolic Laboratory of the Radboudumc. We're interested in collaboration with academic and industrial partners, either bilateral or as part of multi-partner consortia.
Antibodies are compelling proteins that are essential to the immune system and extremely powerful in biotechnology applications; existing as major players in our defence against external agents (viruses, bacteria, etc.), they are also widely used as tools for research, diagnosis and treatments.
Diversity Plus Panel of BioMAP<sup>®</sup> SystemsBioMAP® Systems
This presentation describes the set of 12 BioMAP® systems that comprise the Diversity Plus panel. The BioMAP® systems platform is a drug discovery technology from BioSeek, LLC.
Historically, genetic toxicology has been comprised of bacterial and cell based in vitro assays such as the Ames assay (a bacterial mutagenicity assay), Micronucleus and Chromosomal Aberration assays (mammalian cytogenetic assays), and Mouse Lymphoma Assay (in vitro mammalian cell gene mutation assay). These were routinely used for safety evaluation and are still part of the standard core battery. The emergence of new technologies has facilitated the development of in vitro methods for safe and effective drug and chemical testing.
This BioReliance® toxicology services webinar will explore alternative models, including 3D skin models that comply with the EC Scientific Committee on Consumer Safety (SCCS) recommendations. It will also discuss how the 3Rs (Replace, Reduce, Refine) Principle advocates the exploration of such alternative methods while achieving required goals.
In this webinar, you will learn:
• About in vitro alternatives to animal toxicity testing in pharma, chemical, tobacco, and personal care products.
• How the 3Rs (Replace, Reduce, Refine) Principle advocates exploring alternative methods without compromising the required goals.
• Alternatives to comply with the 7th Amendment to the EC Cosmetics Directive.
Measuring apoptosis in real time with a new luminescent methodMourad FERHAT, PhD
We developed a homogeneous luminogenic annexin V binding assay to detect apoptosis in real time using a multimode plate reader. The detection reagent has two different annexin V fusion proteins engineered to contain complementing domains of a binary luciferase, a substrate for luciferase and a cell impermeable fluorogenic DNA dye to detect necrotic cells. The method allow real-time monitoring of Cellular apoptosis and necrosis in microwell plates without washing steps with a highly sensitive luminescent signal. The AnnexinV luminescent method is amenable to High throughput and is a good alternative to FACS, low-throughput Annexin V-FITC based method.
Peptide Mass Fingerprinting (PMF) and Isotope Coded Affinity Tags (ICAT)Suresh Antre
Analytical technique for identifying unknown protein. The peptide mass are compared to database containing the theoretical peptide masses of all known protein sequences.
Microorganisms such as bacteria, actinomycetes, and fungi are ubiquitous on our planet. They are widely distributed in soil, water, the human body and other environments. Microorganisms and their activities are of great importance to biogeochemical cycles and to all biological systems. Creative Proteomics provides a one-stop proteomics service from sample collection, protein separation, to protein quantification and bioinformatics analysis. We offer both relative quantification (including iTRAQ, TMT and SILAC) and absolute quantification (such as SRM/MRM and PRM) approaches to help you discover, detect and quantify proteins in a broad array of samples. https://www.creative-proteomics.com/services/proteomics-service.htm
Microorganisms such as bacteria, actinomycetes, and fungi are ubiquitous on our planet. They are widely distributed in soil, water, the human body and other environments. Microorganisms and their activities are of great importance to biogeochemical cycles and to all biological systems. Creative Proteomics provides a one-stop proteomics service from sample collection, protein separation, to protein quantification and bioinformatics analysis. We offer both relative quantification (including iTRAQ, TMT and SILAC) and absolute quantification (such as SRM/MRM and PRM) approaches to help you discover, detect and quantify proteins in a broad array of samples. https://www.creative-proteomics.com/services/proteomics-service.htm
Overview Radboudumc Center for Proteomics, Glycomics and Metabolomics april 2015Alain van Gool
An overview of the proteomics, glycomics and metabolomics expertise and capabilities within the Translational Metabolic Laboratory of the Radboudumc. We're interested in collaboration with academic and industrial partners, either bilateral or as part of multi-partner consortia.
Antibodies are compelling proteins that are essential to the immune system and extremely powerful in biotechnology applications; existing as major players in our defence against external agents (viruses, bacteria, etc.), they are also widely used as tools for research, diagnosis and treatments.
Diversity Plus Panel of BioMAP<sup>®</sup> SystemsBioMAP® Systems
This presentation describes the set of 12 BioMAP® systems that comprise the Diversity Plus panel. The BioMAP® systems platform is a drug discovery technology from BioSeek, LLC.
Historically, genetic toxicology has been comprised of bacterial and cell based in vitro assays such as the Ames assay (a bacterial mutagenicity assay), Micronucleus and Chromosomal Aberration assays (mammalian cytogenetic assays), and Mouse Lymphoma Assay (in vitro mammalian cell gene mutation assay). These were routinely used for safety evaluation and are still part of the standard core battery. The emergence of new technologies has facilitated the development of in vitro methods for safe and effective drug and chemical testing.
This BioReliance® toxicology services webinar will explore alternative models, including 3D skin models that comply with the EC Scientific Committee on Consumer Safety (SCCS) recommendations. It will also discuss how the 3Rs (Replace, Reduce, Refine) Principle advocates the exploration of such alternative methods while achieving required goals.
In this webinar, you will learn:
• About in vitro alternatives to animal toxicity testing in pharma, chemical, tobacco, and personal care products.
• How the 3Rs (Replace, Reduce, Refine) Principle advocates exploring alternative methods without compromising the required goals.
• Alternatives to comply with the 7th Amendment to the EC Cosmetics Directive.
Measuring apoptosis in real time with a new luminescent methodMourad FERHAT, PhD
We developed a homogeneous luminogenic annexin V binding assay to detect apoptosis in real time using a multimode plate reader. The detection reagent has two different annexin V fusion proteins engineered to contain complementing domains of a binary luciferase, a substrate for luciferase and a cell impermeable fluorogenic DNA dye to detect necrotic cells. The method allow real-time monitoring of Cellular apoptosis and necrosis in microwell plates without washing steps with a highly sensitive luminescent signal. The AnnexinV luminescent method is amenable to High throughput and is a good alternative to FACS, low-throughput Annexin V-FITC based method.
Peptide Mass Fingerprinting (PMF) and Isotope Coded Affinity Tags (ICAT)Suresh Antre
Analytical technique for identifying unknown protein. The peptide mass are compared to database containing the theoretical peptide masses of all known protein sequences.
Microorganisms such as bacteria, actinomycetes, and fungi are ubiquitous on our planet. They are widely distributed in soil, water, the human body and other environments. Microorganisms and their activities are of great importance to biogeochemical cycles and to all biological systems. Creative Proteomics provides a one-stop proteomics service from sample collection, protein separation, to protein quantification and bioinformatics analysis. We offer both relative quantification (including iTRAQ, TMT and SILAC) and absolute quantification (such as SRM/MRM and PRM) approaches to help you discover, detect and quantify proteins in a broad array of samples. https://www.creative-proteomics.com/services/proteomics-service.htm
Microorganisms such as bacteria, actinomycetes, and fungi are ubiquitous on our planet. They are widely distributed in soil, water, the human body and other environments. Microorganisms and their activities are of great importance to biogeochemical cycles and to all biological systems. Creative Proteomics provides a one-stop proteomics service from sample collection, protein separation, to protein quantification and bioinformatics analysis. We offer both relative quantification (including iTRAQ, TMT and SILAC) and absolute quantification (such as SRM/MRM and PRM) approaches to help you discover, detect and quantify proteins in a broad array of samples. https://www.creative-proteomics.com/services/proteomics-service.htm
Dear Customer,
Stabicon has helped needs of customers in specific areas of the pharmaceutical Analytical Method Development, Validation and Stability study. As the expansion of our analytical services in new technologies and testing methods to help our clients in characterizing future medicines. We understand biology at each of these levels to advance an integrated view of life processes for Biopharmaceuticals. We realize that future medicines (Biopharma) success requires ability to integrate protein testing capabilities which will provide strategic time and cost advantage for Biopharmaceutical sector.
We would like to share our presentation on Biopharma solution, to download the presentation please click on the below link
https://www.box.net/shared/aboumapp9h9gftunenja/1f7c7a6ef1/rss.xml
If you are interested in our Biopharma services, it will be a good idea to interact with our technical team/ commercial team for further clarification, looking forward to your response.
Thanking you and assuring our best service at all time
Regards
Vijay
Project Director
Stabicon Life Sciences Pvt Ltd
Mobile: +919591974355/080-41714280
www.stabicon.com
Dear Customer,
Stabicon has helped needs of customers in specific areas of the pharmaceutical Analytical Method Development, Validation and Stability study. As the expansion of our analytical services in new technologies and testing methods to help our clients in characterizing future medicines. We understand biology at each of these levels to advance an integrated view of life processes for Biopharmaceuticals. We realize that future medicines (Biopharma) success requires ability to integrate protein testing capabilities which will provide strategic time and cost advantage for Biopharmaceutical sector.
We would like to share our presentation on Biopharma solution, to download the presentation please click on the below link
https://www.box.net/shared/aboumapp9h9gftunenja/1f7c7a6ef1/rss.xml
If you are interested in our Biopharma services, it will be a good idea to interact with our technical team/ commercial team for further clarification, looking forward to your response.
Thanking you and assuring our best service at all time
Regards
Vijay
Director Operations
Stabicon Life Sciences Pvt Ltd
Mobile: +919591974355/080-41714280
www.stabicon.com
Bioinformatics Introduction and Use of BLAST ToolJesminBinti
Hi, I am Jesmin, studying MCSE. I think this file will help you if you want to know the basic information about Bioinformatics and the use of BLAST tool. The BLAST tool is the tool that matches the sequences of DNA,RNA and proteins.
Proteomics is a discipline that analyzes the dynamics of protein components, including expression levels and modification states from a holistic perspective, understands the interactions and connections between proteins, reveals the function of proteins and the laws of cell life, and studies all proteins in cells and their behaviours. Creative Proteomics can provide a comprehensive range of proteomics services to help you better conduct research in the drug discovery process, which include: protein gel and imaging analysis, protein identification, protein quantification, top-down proteomics, peptidomics, post-translational modification analysis, and protein-protein interaction. https://www.creative-proteomics.com/services/protein-gel-and-imaging-analysis.htm
#Stabicon product certification process management strives to improve processes and align the needs of regulatory with client objectives on quality product distribution channel. This gives transparency to the process making it more efficient. It include review and testing process to validate that certified products meet certain standards and comply with a stated designed specification/requirement. Product certification process works towards improving efficiency across the various project stakeholders by helping product evaluation and implement best practices.
Commrcialization is the process of bringing new product to services to market . The broader act of commercializ
Commercialization is the process of bringing new product to services to market. The broader act of commercialization entails production, distribution, marketing, sales, customer support, and other key function critical to achieving the commercial success of the new product or service.
n Ayurveda, withania is widely claimed to have potent aphrodisiac, sedative, rejuvenative and life prolonging properties. It is also used as a general energy-enhancing tonic known as Medharasayana, which means 'that which promotes learning and a good memory' and in geriatric problems.
A growth strategy is an organization's plan for overcoming current and future challenges to realize its goals for expansion. Example of growth strategy goals include increasing market share and revenue, acquiring assets, and improving the organization's products or services.
#FormulationDevelopment is as much an art as it is a science, and it's the people that make the difference. Each of the technologies we work with has application two or more dosage forms and multiple routes of administration. Of course, we have all the analytical and production support needed to advance these products into the clinic. For more details please visit on: http://www.stabicon.com/formulation_Development.php
#Imported or Locally Manufactured #medicines without proper pre & post inspection may be rendered substandard at any point along the #medical supply chain, from the point of manufacture through the point of distribution. Regardless of where along the supply chain substandard #medicines are compromised, they pose serious public health risks. for more details please visit on:http://www.stabicon.com/
#Stabicon has been ambitiously established in 2010. Stabicon has a broad range of capabilities and networks in the industry with over 100+ clients and 450+ completed projects for improving our industry partners' growth & human health benefits. Our decade-old experience includes supporting pharmaceutical companies & Institutes for #dosage conversion, Process # #health quality, Cost efficiency & Turnaround time for their stringent regulatory markets products filing to get market authorization. we are proud to lay a #foundations for prosperous future in prevention and cure segment, future #medicine & #FMCG business.
For more details please visit on:http://www.stabicon.com/
#InvitroStudies are critical to the #drug and #wellness product #development due to their ability to provide a basis for #clinical in vivo studies for predicting best delivery model to take Go/No-Go decision. #Our solution on in vitro analyses can provide proof of concept on delivery dosage form in the early stages or reverse pharmacology #development of the active process, when the selectively and possible interactions of the active process, when the selectivity and possible interactions of the candidate drug towards the desired therapeutic target are established. Our team can provide solution map on case to case basis for your specific requirement . For more details please visit on https://www.stabicon.com/In-Vitro.php
Stabicon Life Sciences’ Innovative Solity 3p™* proprietary product technology platform. It enables a wide set of industry and function specific solutions that allows our customers to elevate productivity and create value.
In vitro studies are critical to drug and wellness product development due to their ability to provide a basis for clinical in vivo studies for predicting best delivery model to take Go/No-Go decision. Our solution on in vitro analyses can provide proof of concept on delivery dosage form in the early stages or reverse pharmacology development of the active process, when the selectivity and possible interactions of the candidate drug towards the desired therapeutic target are established.
Orally Inhaled & Nasal Drug Products: delivery systems, components and specialist services for a growing industry sector” This edition is one in the series of ONdrugDelivery publications.
Nutraceutical product is a food or fortified food product that not only supplements the diet but also assists in treating or
preventing disease (apart from anemia), so provides medical benefits. Nutraceuticals are not tested and regulated to the extent of pharmaceutical drugs…
In Ayurveda, Withania is widely claimed to have potent aphrodisiac, sedative, rejuvenative and life prolonging properties. It is also used as a general energy-enhancing tonic known as Medharasayana, which means ‘that which promotes learning and a good memory’ and in geriatric problems .
Imported or locally manufactured medicine without proper & post inspection may be rendered substandard at any point along the medical supply chain, from the point of manufactured the point of distribution. Regar4dless of where along the supply chain substandard medicines are compromised, they pose serious health risks.
Stabicon specialized in managing product quality process, upgrading and introducing advanced technology into products. We are proud to lay a foundation for prosperous future in prevention and cure segment, future medicine & FMCG business. Established in September 2010. Team experience across from design to concept, lab to commercialization of product and with competent technical experts. Automated process driven and prepared for integration with partner site for transparency and real time access on each application.
We express our gratitude and thank you for being the part of our journey through 10 years. without it would not have been possible. We believe in touching lives by providing good quality of life and wellness. You are the witness to our diversification and venturing into future segments of therapeutic and FMCG.
Stabicon specialized in managing product quality process, upgrading and introducing advanced technology into products. We are proud to lay a foundation for prosperous future in prevention and cure segment, future medicine & FMCG business. Established in September 2010. Team experience spans across from design to concept, lab to commercialization of product and with competent technical experts. Automated Process driven and prepared for integration with partner site for transparency and real time access on each application.
Concept to commercialization document gives an insight into the basic prerequisites of the technical & commercial process being followed at Stabicon. Its key input includes various segments of unmet needs in the Quality Management and address development, Analytical and services and innovation technology. Also a simplified business model along with our concept to completion laboratory design gives Stabicon additional efficiency & viability to our clients.
Ability to work in joint ventures and across divisions, culture and countries. Ability to integrate an understand of IP. Scientific expertise and business strategy. Ability to spur creativity while managing commercially. Expertise in functioning and decision making of regulatory requirement. Human resource skills to transform pharmaceutical segment. Dedicated support systems for web interface with real time information. Quality system must meet regulation in multiple markets plus clients internal standards.
Empowering ACOs: Leveraging Quality Management Tools for MIPS and BeyondHealth Catalyst
Join us as we delve into the crucial realm of quality reporting for MSSP (Medicare Shared Savings Program) Accountable Care Organizations (ACOs).
In this session, we will explore how a robust quality management solution can empower your organization to meet regulatory requirements and improve processes for MIPS reporting and internal quality programs. Learn how our MeasureAble application enables compliance and fosters continuous improvement.
The global radiation oncology market size reached US$ 8.1 Billion in 2023. Looking forward, IMARC Group expects the market to reach US$ 14.5 Billion by 2032, exhibiting a growth rate (CAGR) of 6.5% during 2024-2032.
More Info:- https://www.imarcgroup.com/radiation-oncology-market
The Importance of Community Nursing Care.pdfAD Healthcare
NDIS and Community 24/7 Nursing Care is a specific type of support that may be provided under the NDIS for individuals with complex medical needs who require ongoing nursing care in a community setting, such as their home or a supported accommodation facility.
This document is designed as an introductory to medical students,nursing students,midwives or other healthcare trainees to improve their understanding about how health system in Sri Lanka cares children health.
Rate Controlled Drug Delivery Systems, Activation Modulated Drug Delivery Systems, Mechanically activated, pH activated, Enzyme activated, Osmotic activated Drug Delivery Systems, Feedback regulated Drug Delivery Systems systems are discussed here.
Deep Leg Vein Thrombosis (DVT): Meaning, Causes, Symptoms, Treatment, and Mor...The Lifesciences Magazine
Deep Leg Vein Thrombosis occurs when a blood clot forms in one or more of the deep veins in the legs. These clots can impede blood flow, leading to severe complications.
Navigating Challenges: Mental Health, Legislation, and the Prison System in B...Guillermo Rivera
This conference will delve into the intricate intersections between mental health, legal frameworks, and the prison system in Bolivia. It aims to provide a comprehensive overview of the current challenges faced by mental health professionals working within the legislative and correctional landscapes. Topics of discussion will include the prevalence and impact of mental health issues among the incarcerated population, the effectiveness of existing mental health policies and legislation, and potential reforms to enhance the mental health support system within prisons.
Under Pressure : Kenneth Kruk's StrategyKenneth Kruk
Kenneth Kruk's story of transforming challenges into opportunities by leading successful medical record transitions and bridging scientific knowledge gaps during COVID-19.
Global launch of the Healthy Ageing and Prevention Index 2nd wave – alongside...ILC- UK
The Healthy Ageing and Prevention Index is an online tool created by ILC that ranks countries on six metrics including, life span, health span, work span, income, environmental performance, and happiness. The Index helps us understand how well countries have adapted to longevity and inform decision makers on what must be done to maximise the economic benefits that comes with living well for longer.
Alongside the 77th World Health Assembly in Geneva on 28 May 2024, we launched the second version of our Index, allowing us to track progress and give new insights into what needs to be done to keep populations healthier for longer.
The speakers included:
Professor Orazio Schillaci, Minister of Health, Italy
Dr Hans Groth, Chairman of the Board, World Demographic & Ageing Forum
Professor Ilona Kickbusch, Founder and Chair, Global Health Centre, Geneva Graduate Institute and co-chair, World Health Summit Council
Dr Natasha Azzopardi Muscat, Director, Country Health Policies and Systems Division, World Health Organisation EURO
Dr Marta Lomazzi, Executive Manager, World Federation of Public Health Associations
Dr Shyam Bishen, Head, Centre for Health and Healthcare and Member of the Executive Committee, World Economic Forum
Dr Karin Tegmark Wisell, Director General, Public Health Agency of Sweden
2. Topics Covered :
I. Introduction
II. Protein Basic Mechanism
III. Steps in Protein Production
IV.Identification and characterization technique
V. Monitoring protein synthesis
VI.Data Processing Methodology
2www.stabicon.com bd@stabicon.com
4. Therapeutic small molecule are being dominant for more than 100 years
for treatment then why biologics required for therapeutics ?
Why???
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5. How different they are ?
Product Activity Low–high dose High-low dose
Production Chemical synthesis Living organism
Development Limited Trials Extensive Trials
Regulation Non Specific Specific
Toxicity High Low
Elimination Metabolism Endocytosis
Structural Folding Not Required Required
5www.stabicon.com bd@stabicon.com
6. What are these?
What are these large molecule or biomolecules and how similar are they
to human body?
What makes them so specific and effective?
What is the correlation of large molecule / biomolecule to living
machinery?
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7. Mechanism for Signaling
Several types of molecular
modification are involved in
regulation for a signal transfer
such as : glycosylation,
acetylation,etc
7www.stabicon.com bd@stabicon.com
8. Effect in the body
Small molecules rarely elicit secondary signaling, thus effect prevails
until drug adheres to the target site whereas large molecules always
elicit a secondary signaling hence effect remains even after the drug
is eliminated.
8www.stabicon.com bd@stabicon.com
11. Central dogma
Prokaryotic Cell:
DNA –
(Transcription)
RNA – PROTEIN
(Translation)
Eukaryotic Cell:
DNA – RNA – PROTEIN - PROTEIN MODIFIED
(Post Translation)
11www.stabicon.com bd@stabicon.com
15. Biopharma
Biopharmaceuticals are protein with considerable therapeutic structural
diversity. They tend to be between 100 to 1000 times larger than traditional
small molecule drug .
Such complex proteins can't be produced using conventional chemical
synthesis rather than in a living cell under stringently controlled condition.
15www.stabicon.com bd@stabicon.com
16. How are these designed
16www.stabicon.com bd@stabicon.com
21. Characterization Step
Primary structure - peptide mapping
Glycan analysis
Intact Mass analysis
Amino acid and media analysis
Data processing
21www.stabicon.com bd@stabicon.com
22. How to characterize ?
Large scale screening of proteins, their expression, modification and
interaction by using high-throughput approaches
22www.stabicon.com bd@stabicon.com
23. Characterization Required for
Protein identity (mutant protein)
Protein quantity (Expression)
Protein post-translational modifications (up or down)
Protein structure
Protein-protein interaction
Protein localization
Change in any protein property may cause functional
abnormality and might be relevant to pathogenesis.
Tools
Protein Array
Mass Spectrometry
23www.stabicon.com bd@stabicon.com
24. Why Protein by Mass Spectrometry?
MS can unambiguously identify proteins Gel
separated proteins
Proteins in mixture
Protein: protein association
Identify precise post translational changes
Phosphorylation
N- or C- terminal modification
Many more
24www.stabicon.com bd@stabicon.com
27. MALDI Ionization
Protein or
Peptide
Mass Spectrometer Mass/Charge
(m/z)
Ionization
Solution
Phase
Gas
Phase
Matrix assisted laser desorption ionization (MALDI),
Koichi Tanaka
27www.stabicon.com bd@stabicon.com
28. Data Acquisition from MALDI-MSI
Alanine, peptide in
plasma
m/z = 1474.6
Valine, m/z = 1502.7
Alanine
Valine
28www.stabicon.com bd@stabicon.com
29. Protein or
Peptide
Mass Spectrometer Mass/Charge
(m/z)
Ionization
Solution
Phase
Gas
Phase
ESI Ionization
Electrospray ionization (ESI), John B Fenn
29www.stabicon.com bd@stabicon.com
32. How to identify a single protein by MS?
Mass/Charge
(m/z)
Mass
Digest into many peptides Mass of many
peptides
Peptide mass fingerprinting (PMF)
Mass of many peptide fragments
By
Tandem Mass Spectrometry
Single protein identification
32www.stabicon.com bd@stabicon.com
33. Protein mixture Analysis by LC-MS/ MSy
Protein mixture
Digestion
Peptides
400 800 1200 1600
m/z
MS
MS/MS
10 20 30 min0
HPLC
Database
Searching
LLTTIADAAK
SAGGNYVVFGEAK
EDDVEEAVQAADR
All peptide sequences Identification of many proteins
1 sequencing attempt per 0.5 sec.
3600 sequencing attempts in 30 min.
33www.stabicon.com bd@stabicon.com
34. Protein structural Separation
•An ion in a compact-form has a high mobility, and hence shorter drift time,
•The same ion in a more open conformation has a lower mobility, and hence a
longer drift time
Gate
Detector
Neutral Buffer Gas (-ve force)
Ring Electrodes (Potential Gradient. +ve force)
34www.stabicon.com bd@stabicon.com
39. How to identify a single protein by
MS/MS?
Peptides
Theoretical
Spectrum
Database
searching
m/z
Ionization
MS spectrum MS/MS spectrum
Fragmentation
Protein
digestion
Peptide/protein
identification
m/z m/z
200 400 600 800 1000 1200 m/z
K G A FD E L Q
LIFAGKQLEDGR
b
ions
1: L
2: LI3: LIF
4: LIFA
5: LIFAG
6: LIFAGK
7: LIFAGKQ
8: LIFAGKQL
9: LIFAGKQLE
10:LIFAG
KQLED11:LIFAGKQLEDG
y ions
IFAGKQLEDGR:11
FAGKQLEDGR:10
AGKQLEDGR :9
GKQLEDGR :8
KQLEDGR :7
QLEDGR :6
LEDGR :5
EDGR :4
DGR :3
GR :2
R :1
A GF DEL GLI K Q
Q A
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40. N & C terminal Ions
Selected Peptides (parent ions) are fragmented in the mass of a nebulizing
neutral gas. Energy imparted by collision breaks the covalent bond in
parent bonds.
y & b-type ions series thus generated can give us the sequence of the
peptide
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49. Batch Analysis
Batch 1a Batch 2aB
Batch1b Batch2b
Batch1c Batch2c
difference in proteins
Proteins (to identify and quantify proteins in multiple
samples) How many proteins ?
The choice of
method? How
many samples?
How many variability parameter?
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51. Data processing
Using a software product designed to facilitate MS and LCMS analysis of
biopharmaceutical samples
Intact proteins: Comparison of an entire protein(s) against a well-
characterized standard. Identification of differences, and variants that
require further investigation (some could be contaminants).
Peptide map: Comparison of the peptides resulting from a digested
protein against the peptides from the known standard. Identification of
differences in protein coverage, modifications,…
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52. What software Does
Automates data processing and annotation of experimental
results Produces annotated spectra, chromatograms,
coverage maps and tabular data
Facilitates comparisons between a reference standard and
batches of experimental samples
Outputs include formal reports, figure copy/paste, and tabular
data export Frees users to concentrate on important questions
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56. Results Spectra view (Intensity
filter)
Threshold defined
automatically on the
spectra
T
hreshold value
typed in the table
Filt
er applied on the
results table
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57. Results:
Highlight unique peaks
Unique peaks highlighting
Deglycosylated T022
fragment (analyte only)
Glycosylated T022 fragments (control
only)
Control :BP_094 non-deglycosylated
digested VICAM
Analyte :BP_097 deglycosylated 2h
digested VICAM
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58. Result : Peak match data
comparison analyte/control
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59. Results Peak match data
control (glycosylated)
Percentage of each
glycosylation state in control
Control :BP_079 non-deglycosylated
VICAM Analyte :BP_092
deglycosylated 19h VICAM
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60. Results: Peak match data for
analyte (deglycosylated )
Percentage of each
glycosylation state in analyte
Control :BP_079 non-deglycosylated VICAM
Analyte :BP_092 deglycosylated 19h VICAM
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61. Results: Peak match data
comparison analyte/control
You can add your own
comments
Control :BP_079 non-deglycosylated
VICAM Analyte :BP_092
deglycosylated 19h VICAM
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65. Protein digest Analysis
List of the raw data file
Selected analyte compared with the control
Add or remove analyte
Set the selected analyte as
control
Reprocess the data with another
method
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66. Annotation of the peptides
1:T001
First chain of the protein
Trypsin digestion
First digest product
of the chain
1:T001* Modified form of 1:T001
1:T001-002 Missed cleavage between
1:T001 and 1:T002
1:T001-3:T001
Disulfide bridge between
1:T001 and 3:T001
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71. Results :Peak match data comparison
analyte /control
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72. Results: Peak match data
advanced table
When a mass can correspond to several peptides, the different possibilities can be
seen in the advanced view.
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73. Results: Discrimination between two
assignments
The sequence
corresponding to
the fragment
1:T009* of the LC
gave a better
score than the
sequence of 1:T021
If high energy data are available (acquisition with MSE mode), the
fragmentation data can be used to discriminate several assignment for
the same mass.
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75. Thank you
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Stabicon Life Sciences, #4M-413,1st Floor, H R B R
3rd Block, Kammanahalli Main Road, Bangalore –
560043, Karnataka, India.
Tel: +91 80 41250324
bd@stabicon.com