Learn how new technologies from Waters, the RapiFluor-MS Labeling Reagent and the ACQUITY QDa Mass Detector, enable biopharmaceutical development and QC labs to monitor released N-glycans with complementary fluorescence and mass detection. http://www.waters.com/glycans
Simplifying Chromatographic Methods Transfer: Novel Tools for Replicating You...Waters Corporation
Gain a good understanding on the parameters that impact the successful transfer of an LC method from one instrument to another as well as some of the novel tools (i.e., Arc Multi-flow path technology and gradient SmartStart) that have been created to enable the ACQUITY Arc System to replicate established HPLC methods from previous generations of LC equipment.
Transfer of established gradient reversed-phase methods across both HPLC and UHPLC chromatographic instrumentation requires careful consideration of each instrument’s operating parameters and design. The dwell volume, or the system volume between when the solvents are first mixed and the head of the column, can impact the separation. In addition to the dwell volume, the manner in which the solvents are mixed and the formation of the gradient can also vary from one type of LC system to another. Finally, the manner in which the column and solvent are heated can also effect the separation, specifically whether or not the solvent is passively or actively heated prior to the column, or if the air is static or circulated within the column oven.
To understand the effect of these factors may have on methods transfer, both method conditions and instrument specifications must be factored and evaluated when transferring an LC method from one instrument to another.
See the Whole Picture: Using SV-AUC for Empty/Full AAV Capsid AnalysisMilliporeSigma
Watch this webinar here: https://bit.ly/31ZZM3n
Join this webinar for key insights on using the SV-AUC assay for empty/full analysis of your AAV viral vector. We’ll cover the technical requirements for this assay, data interpretation, and finally how this assay fits into the larger picture of AAV characterization.
Recombinant adeno-associated viruses (AAV) are widely used as gene transfer vectors. However, AAV production generates mixed populations of viral capsids containing either complete viral vector genome (full capsids); partially filled, and those lacking the viral genome (empty capsids). Sedimentation Velocity Analytical Ultracentrifugation (SV-AUC) offers a robust, accurate, and consistent method for characterizing empty/full AAV capsid composition. In this webinar we will review the key technical requirements for performing an AUC assay as well as analysis and data interpretation of the results generated.
In this webinar, you will learn:
• Regulatory expectations for empty/full analysis
• Key technical requirements for running an AUC assay and how to interpret the data from the results generated
• How the AUC assay fits into the larger picture of AAV characterization
Webinar: How to Develop a Regulatory-compliant Continued Process Verification...Merck Life Sciences
Participate in the interactive webinar now: http://bit.ly/CPVWebinar
Product life cycle consists of 3 phases: Process Design, Process Performance Qualification and the last and the lengthiest Continued Process Verification (CPV). As more and more biomanufacturing processes enter commercial phases, the critical need to understand how to efficiently perform CPV programs arises.
Explore our webinar library: www.merckmillipore.com/webinars
Biopharmaceutical Attribute Monitoring with the Waters ACQUITY QDa Mass DetectorWaters Corporation
Bringing greater sensitivity, selectivity, and productivity to routine analysis of biotherapeutics, whether you're in characterization or in downstream production of biologics.
Fast, selective, and sensitive methods can be developed for the analysis of impurities
Offering many business benefits using UPLC and UPC2
Increase in sample throughput
Reduction in toxic solvent usage
Using mass spectral detection over UV detection provides
Improvement in sensitivity and selectivity
Reduced matrix effects
PDA and mass detection provide complementary information for peak assignment and structural confirmation of impurities
Adding Mass Detection to Monitor Peptides in Biopharmaceutical Development & QCWaters Corporation
Learn how the Waters ACQUITY QDa Detector is a powerful tool for mass detection in monitoring peptides in HPLC or UPLC assays, in biopharmaceutical late development and quality control. http://www.waters.com/qdabiopharm
This document provides an agenda for a seminar on 5G physical layer technologies. It introduces 5G and compares it to 3G and 4G. It discusses OFDMA, MIMO, waveforms, numerology and frame structure, initial access and beam management, and bandwidth parts. The introduction gives an overview of 5G requirements and the OSI reference model. Later sections provide more details on these topics and their significance for 5G.
Simplifying Chromatographic Methods Transfer: Novel Tools for Replicating You...Waters Corporation
Gain a good understanding on the parameters that impact the successful transfer of an LC method from one instrument to another as well as some of the novel tools (i.e., Arc Multi-flow path technology and gradient SmartStart) that have been created to enable the ACQUITY Arc System to replicate established HPLC methods from previous generations of LC equipment.
Transfer of established gradient reversed-phase methods across both HPLC and UHPLC chromatographic instrumentation requires careful consideration of each instrument’s operating parameters and design. The dwell volume, or the system volume between when the solvents are first mixed and the head of the column, can impact the separation. In addition to the dwell volume, the manner in which the solvents are mixed and the formation of the gradient can also vary from one type of LC system to another. Finally, the manner in which the column and solvent are heated can also effect the separation, specifically whether or not the solvent is passively or actively heated prior to the column, or if the air is static or circulated within the column oven.
To understand the effect of these factors may have on methods transfer, both method conditions and instrument specifications must be factored and evaluated when transferring an LC method from one instrument to another.
See the Whole Picture: Using SV-AUC for Empty/Full AAV Capsid AnalysisMilliporeSigma
Watch this webinar here: https://bit.ly/31ZZM3n
Join this webinar for key insights on using the SV-AUC assay for empty/full analysis of your AAV viral vector. We’ll cover the technical requirements for this assay, data interpretation, and finally how this assay fits into the larger picture of AAV characterization.
Recombinant adeno-associated viruses (AAV) are widely used as gene transfer vectors. However, AAV production generates mixed populations of viral capsids containing either complete viral vector genome (full capsids); partially filled, and those lacking the viral genome (empty capsids). Sedimentation Velocity Analytical Ultracentrifugation (SV-AUC) offers a robust, accurate, and consistent method for characterizing empty/full AAV capsid composition. In this webinar we will review the key technical requirements for performing an AUC assay as well as analysis and data interpretation of the results generated.
In this webinar, you will learn:
• Regulatory expectations for empty/full analysis
• Key technical requirements for running an AUC assay and how to interpret the data from the results generated
• How the AUC assay fits into the larger picture of AAV characterization
Webinar: How to Develop a Regulatory-compliant Continued Process Verification...Merck Life Sciences
Participate in the interactive webinar now: http://bit.ly/CPVWebinar
Product life cycle consists of 3 phases: Process Design, Process Performance Qualification and the last and the lengthiest Continued Process Verification (CPV). As more and more biomanufacturing processes enter commercial phases, the critical need to understand how to efficiently perform CPV programs arises.
Explore our webinar library: www.merckmillipore.com/webinars
Biopharmaceutical Attribute Monitoring with the Waters ACQUITY QDa Mass DetectorWaters Corporation
Bringing greater sensitivity, selectivity, and productivity to routine analysis of biotherapeutics, whether you're in characterization or in downstream production of biologics.
Fast, selective, and sensitive methods can be developed for the analysis of impurities
Offering many business benefits using UPLC and UPC2
Increase in sample throughput
Reduction in toxic solvent usage
Using mass spectral detection over UV detection provides
Improvement in sensitivity and selectivity
Reduced matrix effects
PDA and mass detection provide complementary information for peak assignment and structural confirmation of impurities
Adding Mass Detection to Monitor Peptides in Biopharmaceutical Development & QCWaters Corporation
Learn how the Waters ACQUITY QDa Detector is a powerful tool for mass detection in monitoring peptides in HPLC or UPLC assays, in biopharmaceutical late development and quality control. http://www.waters.com/qdabiopharm
This document provides an agenda for a seminar on 5G physical layer technologies. It introduces 5G and compares it to 3G and 4G. It discusses OFDMA, MIMO, waveforms, numerology and frame structure, initial access and beam management, and bandwidth parts. The introduction gives an overview of 5G requirements and the OSI reference model. Later sections provide more details on these topics and their significance for 5G.
This document discusses Ultra Performance Liquid Chromatography (UPLC). UPLC uses smaller particle sizes of 1.7μm in columns compared to 4μm in HPLC, which allows for faster separations, higher resolution, and shorter run times. UPLC provides improvements in speed, sensitivity and resolution compared to HPLC. It also uses higher pressures up to 15,000 psi compared to HPLC. The document compares various parameters of HPLC and UPLC and discusses their instrumentation including columns, pumps, detectors and applications in pharmaceutical analysis.
The document discusses key principles and guidelines for regulated bioanalysis including validation of quantitative bioanalytical methods. It covers validation of both non-chromatographic and chromatographic assays. Some of the main points covered include validation criteria for calibration curves, quality controls, selectivity, accuracy, precision, reproducibility, recovery, and stability. Examples of validation results are also provided to illustrate concepts like matrix effects, column ruggedness, and recovery.
The document provides information about the Organization for Economic Co-operation and Development (OECD). It discusses the history and establishment of the OECD, its objectives to promote policies that improve economic and social well-being, and how it provides a forum for governments to work together on issues like economic growth, employment, financial stability, and trade. The document also summarizes OECD's toxicity testing guidelines, which provide standardized methods for testing chemicals and their potential hazards. This allows data to be shared between countries and avoids duplicative testing.
This document provides an overview of 3GPP LTE technology. It discusses the evolution of 3GPP standards and the advancement needed for high data rates, including the use of OFDM(A) and SC-FDMA. It provides a brief introduction to LTE including its radio interface architecture, downlink and uplink transmissions, and cell search procedure. Relevant 3GPP specifications for LTE are also listed.
The document introduces key performance indicators (KPIs) for 5G RAN2.0 including service integrity KPIs, utilization KPIs, availability KPIs, and traffic KPIs. Service integrity KPIs measure end user quality and include user and cell throughput. Utilization KPIs evaluate resource usage, such as resource block utilization rate and CPU load. Availability KPIs assess network availability using the radio network unavailability rate. Traffic KPIs quantify uplink/downlink volumes and average/maximum user numbers. Counters are associated with each KPI to calculate values based on network measurements.
Bioanalytical method development and validation .Shubham Bora
1) A bioanalytical method was developed and validated for the quantification of levodopa and carbidopa in rat plasma using LC-MS/MS. Derivatization and ion-pairing chromatography were used to improve the chromatographic retention of the polar analytes.
2) The method was fully validated as per FDA guidelines and demonstrated selectivity, linearity, accuracy, precision, recovery, matrix effects and stability in accordance with acceptance criteria.
3) The validated method was successfully applied to support toxicokinetic studies of levodopa and carbidopa in rats.
Stability indicating RP-HPLC method for estimation of dapagliflozin in bulk a...SriramNagarajan19
A simple, specific, accurate, precise and stability-indicating reverse phase high performance liquid chromatography (RP-HPLC) method is developed for estimation of Dapagliflozin (DGF) in bulk and Pharmaceutical dosage form. The method employed, Hypersil BDS C18 250 mm x 4.6 mm, 5 mm column in isocratic mode with mobile phase of 0.1% Ortho phosphoric acid buffer and acetonitrile 50:50% v/v. The flow rate was 1.0 mL min-1 and effluent was monitored at 245 nm using PDA detector. The injection volume was 10 µl and the total runtime was set as 5min. The retention time for DGF was found to be 2.226min.The method was validated in terms of Linearity, accuracy, precision, limit of detection (LOD), limit of quantification (LOQ) etc. in accordance with ICH guidelines. Linear regression analysis data for the calibration plot showed that there was a good linear relationship between response and concentration in the range of 25 - 150 µg/ml respectively. The LOD and LOQ values for HPLC method were found to be 0.04 and 0.121 µg/ml respectively. No chromatographic interference from the tablet excipients was found. The proposed method was successfully used for estimation of Dapagliflozin (DGF) in Bulk and Pharmaceutical dosage form.
Target Validation / Biochemical and Cellular Assay Development OSUCCC - James
Target validation and assay development are essential steps in the drug discovery process. This document discusses several approaches to target validation, including using genetic tools like CRISPR/Cas9 and RNAi to interrogate targets. It also provides an example of developing a cellular assay using patient-derived cells to validate a target for cystic fibrosis. Additionally, the document describes a case study where phenotypic screening was used to discover a small molecule that restores function of a mutant protein associated with Usher Syndrome type III.
Compendial requirements for particle testing 2014D Scott Aldrich
This document discusses particle counting and identification strategies, outlining compendial guidance from the US, EU, and Japan. It defines particulate matter and describes size ranges and categories. Visible and subvisible particles are monitored using techniques like light obscuration and membrane microscopy. Harmonized limits for subvisible particles are outlined in USP <788>. The document stresses that particle investigation requires a holistic approach using various characterization methods.
This document discusses resveratrol, a natural compound found in red wine and other foods. It provides background on resveratrol chemistry and properties, including its structure as a stilbenoid with three hydroxyl groups. Synthetic methods for producing resveratrol are summarized, including a new efficient synthesis using a Heck reaction. The health benefits of resveratrol are outlined, such as anti-aging and heart health effects. Spectroscopy data is presented to characterize the structure of resveratrol and intermediates produced during synthesis.
This document provides guidance on developing and optimizing a regulated bioanalytical method using liquid chromatography-tandem mass spectrometry (LC-MS/MS). It discusses important considerations for method development including choosing a detection technique, optimizing sample extraction and chromatography conditions, and validating the final method. The goal is to develop a selective, sensitive and reproducible method for quantifying biological samples in a regulated setting.
The feature of the new general chapter 621 is that a column packed with small particles can be used if column length and particle ratio (L/dp) is kept constant between the designated and modified column. This enables high speed analysis of USP methods more than ever.
In this study, a USP method was successfully transferred to an ultra-high speed method with the system suitability requirements met.
For more information, go to SSI.Shimadzu.com. Thanks for viewing.
This presentation correlates the requirements of Annex 11 guidelines to other official regulations and guidance documents.
The correlation is organized in a tabular format.
In the row lists the contents of Annex 11 together with the paragraph numbers.
Rest of the rows correlate the section numbers of
Annex 11 Versions 1993
US FDA 21 CFR Part 211
US FDA Part 820 and
US FDA 21 CFR Part 11
The CMC Journey in the Regulation of Biologicsenarke
Journey in the Development of Biologics Through End of Phase 3
Our Goals
To better understand the FDA’s CMC requirements and expectations for biologic manufacturing and product testing
To better visualize a cost-effective, risk-managed approach to manage these manufacturing processes and products through clinical development into market approval
To better appreciate the challenges involved with controlling safety, potency, and impurity profiles for these products
This document provides an overview and summary of ICH Q10, which describes a model quality management system for the pharmaceutical industry. The objectives of ICH Q10 are to achieve product realization, establish and maintain a state of control, and facilitate continual improvement. It applies throughout the product lifecycle and augments regional GMP requirements. Key elements include process performance and product quality monitoring, corrective and preventive action systems, change management, management review, knowledge management, and quality metrics.
An integrative solution towards SOTIF and AV safetyBernhard Kaiser
Slide set from this year's SOTIF conference in Austin, Texas, Oct 1 and 2, 2019. Shows intermediate pragmatic ideas on how to handle SOTIF in combination with ISO 26262 safety, and how to integrate SOTIF analysis with simulation and driving verification. Terminology may still change as ISO 21448 is evolving.
This document provides an overview of the Enhanced Fast Dormancy feature for WCDMA RAN networks. It describes the technical principles and operation of the feature, including state transitions for fast dormancy user equipment. The document also provides network impact information and engineering guidelines for deploying, monitoring, and optimizing the feature.
Previously certain classes of active substances were required to be manufactured in dedicated or segregated self-contained facilities Certain antibiotics, Certain hormones, Certain cytotoxic ,Certain highly active drugs .This was due to the perceived risk of these active substances.
Pharmaceuticals not covered under these criteria were addressed by a cleaning validation process This involved reduction of the concentration of residual active substance to a level where the maximum carryover from the total equipment train would result in no greater than 1/1000th of the lowest clinical dose of the contaminating substance in the maximum daily dosage of the next product to be manufactured.
This criterion was applied concurrently with a maximum permitted contamination of 10 ppm of the previous active substance in the next product manufactured. Whichever of these criteria resulted in the lowest carryover, constituted the limit applied for cleaning validation. However, these limits did not take account of the available pharmacological and toxicological data They may have been too restrictive or not restrictive enough. EMA therefore felt for a more scientific case by case approach for all classes of pharmaceutical substances.
An Introduction to Self-Organizing Networks (SON)eXplanoTech
This document provides an overview of Self-Organizing Networks (SON) and their key features and functions. It discusses three main aspects of SON: self-configuration, self-optimization, and self-healing. Specific SON techniques covered include automatic neighbor relations, mobility load balancing, mobility robustness optimization, coverage and capacity optimization, and minimization of drive testing. The document also outlines the SON architecture and features defined in 3GPP Releases 8-10.
This presentation describes development of a routine tandem quadrupole LC/MS/MS method for milk and egg allergens based on proteomic studies to identify the allergenic peptide markers. Initial studies are done using a proteomic workflow and quadrupole time of flight mass spectrometry.
From WCBP 2015: GlycoWorks RapiFluor-MS for Glycan ProfilingWaters Corporation
In this vendor presentation given at WCBP, we introduce a new glycan fluorescent label, RapiFluor-MS, which is used to label N-linked glycans. This innovative label improves FLR and mass spectrometry signals for glycan characterization and profiling analysis. Plus - our GlycoWorks RapiFluor-MS N-Glycan Kit now allows you to finish glycan deglycosylation, labeling and cleanup in 3 steps and just 30 minutes.
Learn more about this novel technology and its applications for glycosylated proteins:
http://www.waters.com/glycans
See more of Waters' solutions for biopharmaceutical laboratories:
http://www.waters.com/biopharmaceutical
Playback a full video of this talk, recorded Jan. 27, 2015 at WCBP:
http://www.waters.com/waters/library.htm?cid=10116552&lid=134833758
This document discusses Ultra Performance Liquid Chromatography (UPLC). UPLC uses smaller particle sizes of 1.7μm in columns compared to 4μm in HPLC, which allows for faster separations, higher resolution, and shorter run times. UPLC provides improvements in speed, sensitivity and resolution compared to HPLC. It also uses higher pressures up to 15,000 psi compared to HPLC. The document compares various parameters of HPLC and UPLC and discusses their instrumentation including columns, pumps, detectors and applications in pharmaceutical analysis.
The document discusses key principles and guidelines for regulated bioanalysis including validation of quantitative bioanalytical methods. It covers validation of both non-chromatographic and chromatographic assays. Some of the main points covered include validation criteria for calibration curves, quality controls, selectivity, accuracy, precision, reproducibility, recovery, and stability. Examples of validation results are also provided to illustrate concepts like matrix effects, column ruggedness, and recovery.
The document provides information about the Organization for Economic Co-operation and Development (OECD). It discusses the history and establishment of the OECD, its objectives to promote policies that improve economic and social well-being, and how it provides a forum for governments to work together on issues like economic growth, employment, financial stability, and trade. The document also summarizes OECD's toxicity testing guidelines, which provide standardized methods for testing chemicals and their potential hazards. This allows data to be shared between countries and avoids duplicative testing.
This document provides an overview of 3GPP LTE technology. It discusses the evolution of 3GPP standards and the advancement needed for high data rates, including the use of OFDM(A) and SC-FDMA. It provides a brief introduction to LTE including its radio interface architecture, downlink and uplink transmissions, and cell search procedure. Relevant 3GPP specifications for LTE are also listed.
The document introduces key performance indicators (KPIs) for 5G RAN2.0 including service integrity KPIs, utilization KPIs, availability KPIs, and traffic KPIs. Service integrity KPIs measure end user quality and include user and cell throughput. Utilization KPIs evaluate resource usage, such as resource block utilization rate and CPU load. Availability KPIs assess network availability using the radio network unavailability rate. Traffic KPIs quantify uplink/downlink volumes and average/maximum user numbers. Counters are associated with each KPI to calculate values based on network measurements.
Bioanalytical method development and validation .Shubham Bora
1) A bioanalytical method was developed and validated for the quantification of levodopa and carbidopa in rat plasma using LC-MS/MS. Derivatization and ion-pairing chromatography were used to improve the chromatographic retention of the polar analytes.
2) The method was fully validated as per FDA guidelines and demonstrated selectivity, linearity, accuracy, precision, recovery, matrix effects and stability in accordance with acceptance criteria.
3) The validated method was successfully applied to support toxicokinetic studies of levodopa and carbidopa in rats.
Stability indicating RP-HPLC method for estimation of dapagliflozin in bulk a...SriramNagarajan19
A simple, specific, accurate, precise and stability-indicating reverse phase high performance liquid chromatography (RP-HPLC) method is developed for estimation of Dapagliflozin (DGF) in bulk and Pharmaceutical dosage form. The method employed, Hypersil BDS C18 250 mm x 4.6 mm, 5 mm column in isocratic mode with mobile phase of 0.1% Ortho phosphoric acid buffer and acetonitrile 50:50% v/v. The flow rate was 1.0 mL min-1 and effluent was monitored at 245 nm using PDA detector. The injection volume was 10 µl and the total runtime was set as 5min. The retention time for DGF was found to be 2.226min.The method was validated in terms of Linearity, accuracy, precision, limit of detection (LOD), limit of quantification (LOQ) etc. in accordance with ICH guidelines. Linear regression analysis data for the calibration plot showed that there was a good linear relationship between response and concentration in the range of 25 - 150 µg/ml respectively. The LOD and LOQ values for HPLC method were found to be 0.04 and 0.121 µg/ml respectively. No chromatographic interference from the tablet excipients was found. The proposed method was successfully used for estimation of Dapagliflozin (DGF) in Bulk and Pharmaceutical dosage form.
Target Validation / Biochemical and Cellular Assay Development OSUCCC - James
Target validation and assay development are essential steps in the drug discovery process. This document discusses several approaches to target validation, including using genetic tools like CRISPR/Cas9 and RNAi to interrogate targets. It also provides an example of developing a cellular assay using patient-derived cells to validate a target for cystic fibrosis. Additionally, the document describes a case study where phenotypic screening was used to discover a small molecule that restores function of a mutant protein associated with Usher Syndrome type III.
Compendial requirements for particle testing 2014D Scott Aldrich
This document discusses particle counting and identification strategies, outlining compendial guidance from the US, EU, and Japan. It defines particulate matter and describes size ranges and categories. Visible and subvisible particles are monitored using techniques like light obscuration and membrane microscopy. Harmonized limits for subvisible particles are outlined in USP <788>. The document stresses that particle investigation requires a holistic approach using various characterization methods.
This document discusses resveratrol, a natural compound found in red wine and other foods. It provides background on resveratrol chemistry and properties, including its structure as a stilbenoid with three hydroxyl groups. Synthetic methods for producing resveratrol are summarized, including a new efficient synthesis using a Heck reaction. The health benefits of resveratrol are outlined, such as anti-aging and heart health effects. Spectroscopy data is presented to characterize the structure of resveratrol and intermediates produced during synthesis.
This document provides guidance on developing and optimizing a regulated bioanalytical method using liquid chromatography-tandem mass spectrometry (LC-MS/MS). It discusses important considerations for method development including choosing a detection technique, optimizing sample extraction and chromatography conditions, and validating the final method. The goal is to develop a selective, sensitive and reproducible method for quantifying biological samples in a regulated setting.
The feature of the new general chapter 621 is that a column packed with small particles can be used if column length and particle ratio (L/dp) is kept constant between the designated and modified column. This enables high speed analysis of USP methods more than ever.
In this study, a USP method was successfully transferred to an ultra-high speed method with the system suitability requirements met.
For more information, go to SSI.Shimadzu.com. Thanks for viewing.
This presentation correlates the requirements of Annex 11 guidelines to other official regulations and guidance documents.
The correlation is organized in a tabular format.
In the row lists the contents of Annex 11 together with the paragraph numbers.
Rest of the rows correlate the section numbers of
Annex 11 Versions 1993
US FDA 21 CFR Part 211
US FDA Part 820 and
US FDA 21 CFR Part 11
The CMC Journey in the Regulation of Biologicsenarke
Journey in the Development of Biologics Through End of Phase 3
Our Goals
To better understand the FDA’s CMC requirements and expectations for biologic manufacturing and product testing
To better visualize a cost-effective, risk-managed approach to manage these manufacturing processes and products through clinical development into market approval
To better appreciate the challenges involved with controlling safety, potency, and impurity profiles for these products
This document provides an overview and summary of ICH Q10, which describes a model quality management system for the pharmaceutical industry. The objectives of ICH Q10 are to achieve product realization, establish and maintain a state of control, and facilitate continual improvement. It applies throughout the product lifecycle and augments regional GMP requirements. Key elements include process performance and product quality monitoring, corrective and preventive action systems, change management, management review, knowledge management, and quality metrics.
An integrative solution towards SOTIF and AV safetyBernhard Kaiser
Slide set from this year's SOTIF conference in Austin, Texas, Oct 1 and 2, 2019. Shows intermediate pragmatic ideas on how to handle SOTIF in combination with ISO 26262 safety, and how to integrate SOTIF analysis with simulation and driving verification. Terminology may still change as ISO 21448 is evolving.
This document provides an overview of the Enhanced Fast Dormancy feature for WCDMA RAN networks. It describes the technical principles and operation of the feature, including state transitions for fast dormancy user equipment. The document also provides network impact information and engineering guidelines for deploying, monitoring, and optimizing the feature.
Previously certain classes of active substances were required to be manufactured in dedicated or segregated self-contained facilities Certain antibiotics, Certain hormones, Certain cytotoxic ,Certain highly active drugs .This was due to the perceived risk of these active substances.
Pharmaceuticals not covered under these criteria were addressed by a cleaning validation process This involved reduction of the concentration of residual active substance to a level where the maximum carryover from the total equipment train would result in no greater than 1/1000th of the lowest clinical dose of the contaminating substance in the maximum daily dosage of the next product to be manufactured.
This criterion was applied concurrently with a maximum permitted contamination of 10 ppm of the previous active substance in the next product manufactured. Whichever of these criteria resulted in the lowest carryover, constituted the limit applied for cleaning validation. However, these limits did not take account of the available pharmacological and toxicological data They may have been too restrictive or not restrictive enough. EMA therefore felt for a more scientific case by case approach for all classes of pharmaceutical substances.
An Introduction to Self-Organizing Networks (SON)eXplanoTech
This document provides an overview of Self-Organizing Networks (SON) and their key features and functions. It discusses three main aspects of SON: self-configuration, self-optimization, and self-healing. Specific SON techniques covered include automatic neighbor relations, mobility load balancing, mobility robustness optimization, coverage and capacity optimization, and minimization of drive testing. The document also outlines the SON architecture and features defined in 3GPP Releases 8-10.
This presentation describes development of a routine tandem quadrupole LC/MS/MS method for milk and egg allergens based on proteomic studies to identify the allergenic peptide markers. Initial studies are done using a proteomic workflow and quadrupole time of flight mass spectrometry.
From WCBP 2015: GlycoWorks RapiFluor-MS for Glycan ProfilingWaters Corporation
In this vendor presentation given at WCBP, we introduce a new glycan fluorescent label, RapiFluor-MS, which is used to label N-linked glycans. This innovative label improves FLR and mass spectrometry signals for glycan characterization and profiling analysis. Plus - our GlycoWorks RapiFluor-MS N-Glycan Kit now allows you to finish glycan deglycosylation, labeling and cleanup in 3 steps and just 30 minutes.
Learn more about this novel technology and its applications for glycosylated proteins:
http://www.waters.com/glycans
See more of Waters' solutions for biopharmaceutical laboratories:
http://www.waters.com/biopharmaceutical
Playback a full video of this talk, recorded Jan. 27, 2015 at WCBP:
http://www.waters.com/waters/library.htm?cid=10116552&lid=134833758
Analysis of pesticides in food using both LC- and GC-MS/MS, with data and description of Atmospheric Pressure GC, available on the same system as UPLC-MS/MS with rapid changeover.
Environmental analysis can be extremely challenging due to the low detection levels for toxic contaminants specified by legislation, particularly in drinking water, and the complexity of matrices encountered. Consequently highly selective and sensitive detection methods are required. This presentation provides an introduction to tandem quadrupole mass spectrometry and describes the use of high sensitivity tandem quadrupole mass spectrometry for the analysis of various environmental contaminants including pesticides, endocrine disruptors and polyfluorinated compounds such as PFOS.
This presentation describes a new food testing solution that allows mass detection to be accessible for many of the routine analyses found within a food testing lab. The ACQUITY UPLC, QDa detector in combination with the Empower software solution is fit for purpose and is easy to use. You simply power on and you are ready to go
With the ACQUITY QDa detector many of the normal processes that are required for mass spectrometers (such as mass calibration and optimisation and manual adjustments that need to be made) have all been fully automated.
This presentation will provide an introduction to the technology and benefits of Waters ACQUITY Ultra Performance Convergence Chromatography system (UPC2) and illustrate how it can impact the analysis of chemicals and polymers.
Comprehensive Investigation of the Utilization of SFC/ESI Positive Mode MS fo...Waters Corporation
This document discusses the use of single column convergence chromatography/mass spectrometry (UPC2/MS) for bioanalytical studies. It provides examples of how UPC2/MS can simplify workflows by reducing sample preparation times through direct injection of extracts and improving selectivity over reversed phase chromatography. UPC2/MS also allows for faster separation of challenging compound classes like isomers and lipids compared to traditional techniques like gas chromatography. The document concludes that UPC2/MS provides an orthogonal separation method and combines multiple techniques into one analytical platform for streamlining quantitative bioanalysis in drug discovery and development.
Food fraud is on the increase globally and checking for the authenticity of foodstuffs is a major challenge. Typical of food fraud is the substitution of a low value product for a higher value one to increase profit to the trader. For years, traders have been passing off a lesser quality rice, CSR 30, as the world's finest long-grained, aromatic rice, Basmati, in key markets like the US, Canada and the EU. In the process, the rice exports enjoy the duty exemption accorded to pure Basmati in the EU and thousands of consumers get duped. This presentation describes the use of Atmospheric Pressure Gas Chromatography coupled to high resolution MS to differentiate between types of rice and to identify marker compounds using statistical analysis software.
Impact of novel ms ms all acquisition and processing techniques on forensic t...SCIEX
This document evaluates different MS/MS acquisition techniques for forensic toxicological screening using high resolution mass spectrometry. It compares standard data-dependent acquisition to various data-independent acquisition (SWATH) methods. SWATH acquisition ensures MS/MS data is collected for all possible precursors but can result in mixed spectra from multiple compounds. Overlapping variable-width SWATH windows and deconvolution techniques improve spectral quality and identifications over standard wide windows. These methods provide identification results comparable to targeted methods while analyzing all compounds.
This document discusses using Oasis PRiME HLB sorbent for sample cleanup of food matrices prior to multi-residue veterinary drug analysis by LC-MS/MS. Oasis PRiME HLB is a reversed-phase solid phase extraction device that provides a simpler, faster, and cleaner sample preparation compared to other sorbents. It efficiently removes phospholipids and other matrix interferences from complex samples like milk and tissues. Application examples demonstrate excellent recovery of over 60 veterinary drug residues from milk and tissues using a simple pass-through protocol with Oasis PRiME HLB.
Allergens are a major food safety concern and incidences of food allergy in industrialised populations has increased in recent times. One of the most common food allergies is that of peanuts. Food regulations for allergens exist in many countries and are being modified regularly as more is understood about allergens and the reactions they cause. This presentation describes the use of time-of-flight mass spectrometry to locate, identify and quantify an allergenic protein in both raw and roasted peanuts. Typical food processing (e.g. food processing) can alter the markers peptides present and amount that they are present in the samples which adds complexity to the analysis.
Remote sensing –Beyond images
Mexico 14-15 December 2013
The workshop was organized by CIMMYT Global Conservation Agriculture Program (GCAP) and funded by the Bill & Melinda Gates Foundation (BMGF), the Mexican Secretariat of Agriculture, Livestock, Rural Development, Fisheries and Food (SAGARPA), the International Maize and Wheat Improvement Center (CIMMYT), CGIAR Research Program on Maize, the Cereal System Initiative for South Asia (CSISA) and the Sustainable Modernization of the Traditional Agriculture (MasAgro)
The document discusses natural biotoxins and challenges in their analytical screening and confirmation. It provides an overview of common mycotoxins and regulated levels in different food commodities. Current screening methods like immunoassays, TLC and LC-UV/FL are described along with their performance criteria. The benefits of mass detection using ACQUITY QDa for routine multi-toxin screening are highlighted. The document demonstrates a screening method for 12 regulated mycotoxins in wheat using UPLC-MS/MS. It also discusses quantitative confirmatory methods using Xevo TQ-S and validation requirements as per EU legislation.
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The document discusses the benefits of meditation for reducing stress and anxiety. Regular meditation practice can help calm the mind and body by lowering heart rate and blood pressure. Studies have shown that meditating for just 10-20 minutes per day can have significant positive impacts on both mental and physical health over time.
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This document provides information on using the BioPAT Trace system to monitor glucose and lactate concentrations in bioreactor processes. It discusses the measurement principles, which use the enzymes glucose oxidase and lactate oxidase. Data is presented to demonstrate the system's linearity, precision, recovery, operational stability, and shelf life for glucose and lactate sensors. Consumables required for using the system are also listed.
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This presentation discusses the history of size exclusion chromatography from gel permeation to modern day columns, and introduces the next evolution in polymer chromatography, the ACQUITY Advanced Polymer Chromatography system.
Similar to Monitoring Released N-Glycans in Biopharma Development/QC with Fluorescence & Mass Detection (20)
Using Fusion QbD as an Analytical Quality by Design Software for Method Devel...Waters Corporation
This presentation describes the benefits of a hardware and software platform that dramatically advances LC and LC-MS method development by applying Analytical Quality by Design (AQbD) approaches in a 100% regulatory compliance supported framework. This AQbD aligned platform includes Waters Empower™ Chromatography Data System Software with enhanced Fusion QbD® Software, the Waters® ACQUITY UPLC H-Class PLUS, a PDA detector, and QDa Mass Detector. New software capabilities that optimize and simplify the use of mass detection in the AQbD method development workflow have been added.
Visit methods.waters.com for more information
Webinar - Pharmacopeial Modernization: How Will Your Chromatography Workflow ...Waters Corporation
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Or: Beyond linear.
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Disclaimer: No one is perfect, so please mind that there might be mistakes and typos.
dtubbenhauer@gmail.com
Corrected slides: dtubbenhauer.com/talks.html
Hello to all of you and welcome to a presentation about innovative technology solutions from Waters!
We’re really excited to share with you a fast and powerful new solution we have developed for released N-glycan monitoring, and that we think will greatly benefit scientists like you working in biopharmaceutical development, production and QC environments.
Building upon our core ACQUITY UPLC Systems and industry-leading Empower chromatography data software (CDS), the released N-glycan monitoring solution we’re talking about is made possible by two highly innovative new products from Waters – the first being our groundbreaking new GlycoWorks RapiFluor-MS N-Glycan Kit that we launched at the WCBP conference in January (2015), and the second being our ACQUITY QDa Mass Detector, which is another revolutionary product we introduced in the Fall of 2013.
As part of this presentation I’ll take a little time to introduce you to both these products, so you understand what makes them so unique and powerful, and then we’ll focus on how RapiFluor-MS labeling reagent combines with our ACQUITY UPLC H-Class Systems, UPLC glycan columns, and Empower Software to make for a very fast and powerful new solution for N-glycan monitoring on a routine basis.
RapiFluor-MS delivers a 16x faster sample preparation protocol compared to traditional labeling techniques, and it’s 8x faster than other “instant” labeling techniques. In addition, it provides faster chromatographic runs and more accurate results backed up by mass confirmation, and all within a GMP-compliant ready workflow. Sound interesting? Let’s jump in.
To begin, here is a basic outline of the presentation…
We’ll start by first sharing some feedback we’ve been hearing from biopharma customers about the challenges they face today when doing routine glycan analysis. And as our new RapiFluor-MS N-Glycan Kit and the ACQUITY QDa mass detector are key components to our overall solution to address these challenges, we’ll provide a brief introductory overview of both these products.
Next, we’ll look at data that our scientific team has generated, which demonstrate the enhanced speed, performance and productivity that can be achieved when utilizing our new solution. And finally, we’ll summarize what we discussed and demonstrate that putting our new glycan monitoring solution into practice will not only bring you greater speed, but greater confidence in your glycan assignments for better decision making and lower overall risk, and that it can significantly improved productivity as well.
So let’s start with what we’ve been hearing from biopharmaceutical developers and manufacturers like yourself…
The quotes on this slide clearly indicate a level of frustration out there with regard to current released N-glycan methods, especially with regard to the long sample prep time, but also with regard to the data sometimes not providing enough clarity.
So what if you could reduce sample prep time from a day down to just 30 minutes? And what if you could capture both fluorescent data and mass data in a single run, such that you can gain greater confidence in your results with increased sensitivity and mass confirmation? And what if you could do all of this in a GMP compliant manner using your existing workflow, and you could speed up sample runs by 50% or more?
Well, that’s just what you can do with our new released N-glycan monitoring solution and I’ll show you data that backs this up. But first let me take a few moments to introduce you to the two products that make this new solution possible: our brand new RapiFluor-MS N-Glycan Kit and our ACQUITY QDa detector.
The key enabler for our new solution to N-glycan monitoring is our recently launched RapiFluor-MS reagent.
To develop this reagent, we revisited Waters’ expertise in rapid, fluorescence labeling of amino acids
For 20 years, Waters has been providing a solution for amino acid labeling that uses the novel reagent, known as AccQ•Fluor.
This molecule has 3 very important chemical characteristics;
It has a rapid tagging reactive group, and
It has an efficient fluorophore for high sensitivity fluorescence detection, and
To facilitate modern N-glycan analyses, AccQ Fluor was functionalized so as to have a third chemical property, an MS enhancing tertiary amine charge tag. This has produced a novel labeling reagent that we call RapiFluor-MS.
Beyond our new label, the GlycoWorks RapiFluor-MS N-Glycan Kit also includes a new and proprietary enzyme mixture which greatly simplifies the deglycosolation process and reduces deglycosolation time from 16+ hours down to less than 15 minutes!!! Yes - you heard that correctly.
In fact, with the RapiFluor-MS N-Glycan Kit you can complete your entire sample prep in just 30 minutes! This remarkable improvement not only enhances productivity on the sample prep side, but also on the detection side, by enabling more rapid methods (explained later in the presentation) and by enabling both fluorescent detection and mass detection within a single GMP compliant workflow, thereby increasing certainty especially with overlapping peaks, and reducing the amount of follow-on analysis that may be needed.
Comparing FLR and MS sensitivity to 3 typical labels used on the market. FLR is in red, and MS in blue.
Because the RapiFluor-MS label has high proton affinity, derivatized glycans preferentially adopt uniquely high charge states during positive ion mode electrospray ionization. As shown with the mass spectra on the left, the predominant charge state for a small neutral glycan, such as an FA2 glycan, labeled with RapiFluorMS is 2+, although it increases to 3+ for larger molecular weight glycans, such as an FA2BG2S2, disialylated glycan.
Notice that these species when labeled with 2-AB adopt only 1+ and 2+ charge states, respectively.
Now let me take a few moments to tell you about the ACQUITY QDa detector, another pioneering product that exemplifies our innovation strategy here at Waters. Launched in 2013, it has seen remarkable uptake across multiple industries and is considered one of the most successful product introductions in the company’s history, which is saying a lot!
Our goal from the outset was to allow analytical chemists to collect mass information in a similar way to how they currently collect optical information, by providing a compact, robust, and highly affordable mass detector that’s as easy to use as a PDA detector, and that seamlessly integrates into existing Empower based UPLC systems. That is exactly what we did.
The instrument control, data acquisition and reporting functions are all fully integrated into our industry leading Empower Chromatography Data Software (Empower 2 FR5 and Empower 3), so anyone using an Empower based UPLC system today can literally add this unit to their existing stack and begin collecting mass data right away.
Here you see how the ACQUITY QDa Detector is designed to slot right into an existing stack. It is easily added to all existing ACQUITY UPLC systems running with Empower 2 (FR5) or Empower 3, which enables the data to be captured and reported in a GMP compliant manner. The QDa has been designed to work with both 110 and 220V power, so importantly in North America it does not require any special electrical hook-up. It also requires minimal maintenance, and requires minimal training to operate, all of which makes it incredibly easy to implement.
(Note: GUI = Graphical User Interface)
As previously indicated, adding an ACQUITY QDa to an existing stack and running it is no more difficult than that of a PDA detector. In fact the GUI interfaces within Empower Software for the set-up, data viewing and reporting are all identical to that of a FLR detector. The interface on this slide shows how users program the instrument to collect mass data. Users can easily select the mass range to be scanned, what ion detection mode to use (positive, negative or both), and through the advanced tab, one can easily create a list of target analytes they wish to monitor by using selected ion recording (SIR).
The ACQUITY QDa performs automated calibration and resolution with every start-up, ensuring data collected is accurate and precise every time the instrument is used. In addition, the electrospray (ESI) interface has been optimized to preserve the resolution of your separation. The pre optimized source greatly limits the amount of “tuning” necessary to get good results, a factor which has been and continues to be a key challenge when introducing mass spectrometry to new settings. With the ACQUITY QDa, we have addressed these issues and made it much easier for chromatographers and analytical chemists to start generating their own mass data.
As mentioned previously, the ACQUITY QDa is incredibly robust and requires minimal maintenance, which is often not the case with mass spectrometry equipment. There are just two components that periodically need to be replaced - the sample aperture, and the capillary assembly - and to make replacement as easy as possible to maintain, we designed these parts to be disposable.
On the left you can see how it is to remove the sample aperture. Once removed it is discarded and replaced with a new aperture, and the entire process can be completed in under 10 minutes.
On the right is a pre-cut and assembled capillary assembly, which can easily be replaced in under a minute. S No cutting or assembly required on the part of the end user.
So in summary, the ACQUITY QDa is yet another pioneering, innovative product from Waters that has experienced tremendous success since it’s market introduction.
As simple to deploy as a PDA, easy to use and maintain, highly compact and affordable – the ACQUITY QDa is a groundbreaking product that is expanding and strengthening the capabilities of analytical chemists by allowing them to collect mass data routinely for greater confidence and insight in their separations, and that is fueling productivity gains in many industrial workflows, including those of pharmaceutical and biopharmaceutical development, production and QC.
So now that you know more about these two innovative products, let me walk you through some released N-glycan data we recently generated, which shows:
how well the QDa detects RapiFluor-MS labeled N-glycans;
how the QDa fully integrates with our industry leading Empower CDS software, making it a very simple add-on to your existing chromatographic workflow – allowing you to capture, annotate and report mass data in a GMP compliant manner; and
how the QDa and RapiFluor-MS combination enables much faster and higher throughput N-glycan profiling on a routine basis, which can speed up process development and optimization and improve productivity overall.
So let’s look first at N-glycan detection with the RapiFluor-MS label using both a fluorescence detector and the QDa mass detector. Represented on this slide are the combined chromatograms of released N-glycans from three different samples chosen to reflect the wide range of glycans typically seen and monitored in biotherapeutic development and production settings. These are 1) IgG, which includes simple bi-antennary glycan structures, 2) Rnase B, which includes many high mannose structures, and 3) Bovine Fetuin, which includes many larger, more complex and highly sialylated glycan structures. As you can see, both chromatograms show equivalent detection responses, and all the peaks in one are seen in the other.
The FLR trace is on top, and the TIC trace is on the bottom.
Analytical chemists now have the ability to detect and monitor glycoforms not just by retention time and fluorescent signal, but my mass data as well, and that all of this data can be captured simultaneously in a single run, simply by adding the ACQUITY QDa as an orthogonal mass detector to existing Empower based chromatographic workflows.
On this slide we show the ability of the QDa to detect RapiFluor-MS labeled N-glycans across a wide dynamic range. Shown on the right side are the mass spectra for the two peaks that represent the most abundant and least abundant glycoforms in this IgG glycan profile (FA2 and A2G1b respectively).
Note, RPA% is calculated based on peak areas from the fluorescent trace.
On this slide we list all of the released N-glycans from an IgG MS standard, and the charge state by which the QDa detects each glycoform.
For a more complex set of glycoforms we looked at Bovine Fetuin glycans, which include some the largest, most complex and highly sialylated glycoforms that can be associated with protein therapeutics. As shown, all peaks on the FLR trace are seen with equivalent response levels on the TIC trace using the QDa - further demonstrating the ability of the QDa to accurately detect large and complex glycoforms tagged with the RapiFluor-MS label, and underscoring its utility as a robust glycan monitoring tool.
Here we look at the mass spectra for one of the largest and most complex glycoforms in the Fetuin profile. The A3G3S4 glycan is a large tri-antennary tetra-sialylated glycoform, and as shown, its +3 charge state clearly appears within the ACQUITY QDa mass window.
To determine if other charge states are perhaps not being seeing with the ACQUITY QDa, we analyzed the same glycoform using our Xevo G2-XS QTof mass spectrometer, which confirmed only the +3 charge state exists for this glycoform.
With the QDa fully integrated into Empower, both optical and mass data can now be captured within a single run, and as shown on this slide, it’s easy to annotate fluorescent chromatograms with peak names and corresponding mass data.
Empower is the leading Chromatography Data Software (CDS) in the industry because of it’s ease-of-use, intuitive functionality, and its comprehensive GMP compliant ready data analysis and reporting capabilities. And with it now being fully integrated with the QDa, all the mass data captured can be combined with optical data and presented in a consolidated GMP compliant report, making it a great solution for biotherapeutic development, production and QC settings.
On this slide we show a 55 minute chromatographic method that has been compressed down to a 10 minute method. You can see there is a loss in chromatographic resolution with the 10 minute method, but if you know what you are looking for and have developed an SIR list (as seen on the next slide), you retain the ability to confirm glycan structures via mass. This can come in handy when doing process development and optimization work and are having to evaluate many different samples/permutations (clone selection, various bioreactor conditions, etc…). The quick 10 minute SIR method in this case enables a higher throughput workflow enabling you to evaluate a wide range of samples more quickly.
Materials and Methods:
C: ACQUITY UPLC BEH Amide column, 1.7 m, 2.1 mm x 50 mm
F: 0.8 mL/min
T: 60 C
A: Acn
B: 50 mM AF, pH 4.5
Gradient for 10 minute method:
T F %ACN
0 0.8 75
6 0.8 54
6.25 0.4 0
7 0.4 0
7.25 0.4 0
8 0.8 75
10 0.8 75
Gradient for 55 minute method:
T F %ACN
0 0.4 75
35 0.4 54
36.5 0.2 0
39.5 0.2 0
43.1 0.2 75
47.6 0.4 75
55 0.4 75
To illustrate this, here we see the 10 minute method with both the fluorescent chromatogram and the SIR overlay, along with the individual SIR’s for each of the targeted glycans being monitored. The key point here is that with the QDa added to your workflow, you not only gain mass confirmation for unambiguous glycan assignments, but you increase productivity as well. With peak identities confirmed via mass, you don’t need as much chromatographic resolution and can run faster methods, thus increasing throughput and accelerating bioprocess development work. (clone selection, bioreactor conditions, etc…)
Increases in Mannose 5 are generally considered undesirable for a host of reasons. Here we simulate an increasing amount of the M5 Mannose structure over time relative to another glycoform: F(6)A2G(4)1. In the table report you can see how the ratio increases with each successive increment in time. This just demonstrates another way the QDa can improve monitoring capabilities and productivity.
To summarize, our new Glycoworks RapiFluor-MS N-glycan kit represents a tremendous breakthrough in released N-glycan analysis and monitoring. Not only are sample prep times cut down from a day and a half to under 30 minutes, but glycan assignments can now be confirmed via orthogonal mass detection within a single workflow. This not only lowers risk by strengthening confidence in glycan assignments, but the use of mass detection also enables much faster and higher throughput methods, which can significantly increase productivity, especially during process development and optimization work.
We’ve shown that the QDa accurately detects a wide range of RapiFluor-MS labeled N-glycans, from simple to highly complex, with excellent sensitivity and selectivity, and across a wide dynamic range. And again, the unique ability to set up SIR lists for monitoring can facilitate higher throughput analysis and strengthen monitoring, especially during process development and optimization.
With RapiFluor-MS and the QDa you can now carry out fast, powerful N-glycan profiling on a routine basis, with unrivaled sensitivity and mass confirmation, all within a single workflow, in your own lab, without having to rely upon external MS resources.
Thank you!
Reimagine glycan analysis with RapiFluor-MS released N-glycan label and the ACQUITY QDa Detector: www.waters.com/glycans
www.waters.com/glycans
More info on the ACQUITY QDA Detector for Biopharm: www.waters.com/qdabiopharm
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Details on the ACQUITY QDa Detector: www.waters.com/qda
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