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Determination of PAHs in Combustion-Related Samples via Multidimensional Chromatographic MethodsGas chromatography–mass spectrometry (GC–MS), reversed-phase LC with stop-flow fluorescence (FL), and constant energy synchronous fluorescence spectroscopy (CESFS) are explored to determine PAH isomers in three combustion-related standard reference materials.
Latest:
Determination of PAHs in Combustion-Related Samples via Multidimensional Chromatographic MethodsGas chromatography–mass spectrometry (GC–MS), reversed-phase LC with stop-flow fluorescence (FL), and constant energy synchronous fluorescence spectroscopy (CESFS) are explored to determine PAH isomers in three combustion-related standard reference materials.
Latest:
Determination of PAHs in Combustion-Related Samples via Multidimensional Chromatographic MethodsGas chromatography–mass spectrometry (GC–MS), reversed-phase LC with stop-flow fluorescence (FL), and constant energy synchronous fluorescence spectroscopy (CESFS) are explored to determine PAH isomers in three combustion-related standard reference materials.
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State-of-the-Art Biopharmaceutical AnalysisThe articles presented here highlight important recent developments in biopharmaceutical analysis.
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Not (Only) Reversed-Phase Liquid Chromatography–Mass Spectrometry: Alternative LC–MS ApproachesReversed-phase LC–MS has limitations in numerous analytical applications. Alternative MS-compatible chromatographic techniques separate analytes in the liquid phase based on different retention mechanisms compared with reversed-phase LC. This article describes these alternative chromatographic approaches, relevant applications, and the future of these techniques.
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Monitoring of On-column Methionine Oxidation as Part of a System Suitability Test During UHPLC–MS/MS Peptide MappingLong-term column use can lead to on-column methionine oxidation during LC–MS/MS peptide mapping of antibody-based biotherapeutics. Following the approach described here minimizes the risk of measuring oxidative artifacts, and helps generate high quality data to provide reliable quantitative information about product-related heterogeneities.
Latest:
Monitoring of On-column Methionine Oxidation as Part of a System Suitability Test During UHPLC–MS/MS Peptide MappingLong-term column use can lead to on-column methionine oxidation during LC–MS/MS peptide mapping of antibody-based biotherapeutics. Following the approach described here minimizes the risk of measuring oxidative artifacts, and helps generate high quality data to provide reliable quantitative information about product-related heterogeneities.
Latest:
Monitoring of On-column Methionine Oxidation as Part of a System Suitability Test During UHPLC–MS/MS Peptide MappingLong-term column use can lead to on-column methionine oxidation during LC–MS/MS peptide mapping of antibody-based biotherapeutics. Following the approach described here minimizes the risk of measuring oxidative artifacts, and helps generate high quality data to provide reliable quantitative information about product-related heterogeneities.
Latest:
Monitoring of On-column Methionine Oxidation as Part of a System Suitability Test During UHPLC–MS/MS Peptide MappingLong-term column use can lead to on-column methionine oxidation during LC–MS/MS peptide mapping of antibody-based biotherapeutics. Following the approach described here minimizes the risk of measuring oxidative artifacts, and helps generate high quality data to provide reliable quantitative information about product-related heterogeneities.
Latest:
Size-Exclusion Chromatography for the Analysis of Complex and Novel Biotherapeutic ProductsThis study presents applications of size-exclusion chromatography (SEC) for characterization and quality control of novel biotherapeutic products, including antibody–drug conjugates, hydrophobic proteins, and coformulations. Examples of modifying SEC mobile-phase composition and running conditions to modulate the separation are discussed, as well as approaches and strategies for analyzing atypical protein products such as coformulations.
Latest:
Size-Exclusion Chromatography for the Analysis of Complex and Novel Biotherapeutic ProductsThis study presents applications of size-exclusion chromatography (SEC) for characterization and quality control of novel biotherapeutic products, including antibody–drug conjugates, hydrophobic proteins, and coformulations. Examples of modifying SEC mobile-phase composition and running conditions to modulate the separation are discussed, as well as approaches and strategies for analyzing atypical protein products such as coformulations.
Latest:
Size-Exclusion Chromatography for the Analysis of Complex and Novel Biotherapeutic ProductsThis study presents applications of size-exclusion chromatography (SEC) for characterization and quality control of novel biotherapeutic products, including antibody–drug conjugates, hydrophobic proteins, and coformulations. Examples of modifying SEC mobile-phase composition and running conditions to modulate the separation are discussed, as well as approaches and strategies for analyzing atypical protein products such as coformulations.
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Ferritin as a Natural Protein Scaffold: Building a Multivalent Ferritin–Fab ConjugateIn this study, a conjugation strategy for Fab–ferritin conjugates, used for drug delivery, was successfully optimized using LC–MS. Characterization of the resulting conjugates was performed using SEC-MALS-QELS.
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Ferritin as a Natural Protein Scaffold: Building a Multivalent Ferritin–Fab ConjugateIn this study, a conjugation strategy for Fab–ferritin conjugates, used for drug delivery, was successfully optimized using LC–MS. Characterization of the resulting conjugates was performed using SEC-MALS-QELS.
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Ferritin as a Natural Protein Scaffold: Building a Multivalent Ferritin–Fab ConjugateIn this study, a conjugation strategy for Fab–ferritin conjugates, used for drug delivery, was successfully optimized using LC–MS. Characterization of the resulting conjugates was performed using SEC-MALS-QELS.
Latest:
Ferritin as a Natural Protein Scaffold: Building a Multivalent Ferritin–Fab ConjugateIn this study, a conjugation strategy for Fab–ferritin conjugates, used for drug delivery, was successfully optimized using LC–MS. Characterization of the resulting conjugates was performed using SEC-MALS-QELS.
Latest:
Ferritin as a Natural Protein Scaffold: Building a Multivalent Ferritin–Fab ConjugateIn this study, a conjugation strategy for Fab–ferritin conjugates, used for drug delivery, was successfully optimized using LC–MS. Characterization of the resulting conjugates was performed using SEC-MALS-QELS.
Latest:
Ferritin as a Natural Protein Scaffold: Building a Multivalent Ferritin–Fab ConjugateIn this study, a conjugation strategy for Fab–ferritin conjugates, used for drug delivery, was successfully optimized using LC–MS. Characterization of the resulting conjugates was performed using SEC-MALS-QELS.
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Analytical Applications of Immobilized Enzyme Reactors (IMERs) Coupled to LC–MS/MS for Bottom- and Middle-Up Characterization of ProteinsFlow-through immobilized-enzyme reactors (IMERs) can streamline protein digestion for bottom- and middle-up LC–MS characterization while providing high accuracy and reproducibility.
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Utilizing Multidimensional LC–MS for Hydroxyl Radical Footprinting AnalysisThe potential of multidimensional online peptide mapping analysis as a strategy for improving a postlabeling workflow for protein–protein interactions is demonstrated using both hydroxy radical footprinting–mass spectrometry (HRF–MS) and LC–MS/MS.
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Utilizing Multidimensional LC–MS for Hydroxyl Radical Footprinting AnalysisThe potential of multidimensional online peptide mapping analysis as a strategy for improving a postlabeling workflow for protein–protein interactions is demonstrated using both hydroxy radical footprinting–mass spectrometry (HRF–MS) and LC–MS/MS.
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Development of a Size-Exclusion Chromatography Method to Characterize a Multimeric PEG–Protein ConjugateA simple and robust size-exclusion chromatography (SEC) method has been developed for characterizing a multimeric PEG–protein conjugate. A wide range of size variants of the conjugate, ranging from 50 kDa to >1000 kDa, can be resolved and quantitated.
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Development of a Size-Exclusion Chromatography Method to Characterize a Multimeric PEG–Protein ConjugateA simple and robust size-exclusion chromatography (SEC) method has been developed for characterizing a multimeric PEG–protein conjugate. A wide range of size variants of the conjugate, ranging from 50 kDa to >1000 kDa, can be resolved and quantitated.
Latest:
Development of a Size-Exclusion Chromatography Method to Characterize a Multimeric PEG–Protein ConjugateA simple and robust size-exclusion chromatography (SEC) method has been developed for characterizing a multimeric PEG–protein conjugate. A wide range of size variants of the conjugate, ranging from 50 kDa to >1000 kDa, can be resolved and quantitated.
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Effects of Buffer Capacity in LC, Part II: Visualization of pH Changes Inside the ColumnIn situ measurements of the mobile-phase pH before and after the column help to rationalize the effects of mismatch in pH and concentration between the mobile phase and sample buffer mismatch in reversed-phase LC separations.
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Sample Preparation Goes Subzero: Ice Concentration Linked with Extractive Stirrer (ICECLES)A new technique, ice concentration linked with extractive stirrer (ICECLES), provides highly efficient isolation of trace compounds from aqueous samples. ICECLES combines freeze concentration with stir-bar sorptive extraction into a seamless analytical extraction methodology. This month, we discuss how to perform this relatively simple technique, and discuss its advantages and limitations.
Latest:
Sample Preparation Goes Subzero: Ice Concentration Linked with Extractive Stirrer (ICECLES)A new technique, ice concentration linked with extractive stirrer (ICECLES), provides highly efficient isolation of trace compounds from aqueous samples. ICECLES combines freeze concentration with stir-bar sorptive extraction into a seamless analytical extraction methodology. This month, we discuss how to perform this relatively simple technique, and discuss its advantages and limitations.
Latest:
Sample Preparation Goes Subzero: Ice Concentration Linked with Extractive Stirrer (ICECLES)A new technique, ice concentration linked with extractive stirrer (ICECLES), provides highly efficient isolation of trace compounds from aqueous samples. ICECLES combines freeze concentration with stir-bar sorptive extraction into a seamless analytical extraction methodology. This month, we discuss how to perform this relatively simple technique, and discuss its advantages and limitations.
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A Review of Recent Developments in Analytical Characterization of Glycosylation in Therapeutic ProteinsAdvanced separation and mass spectrometry methods enable comprehensive profiling of the inherent glycan heterogeneities of protein therapeutics. In particular, reversed-phase HPLC–based multiattribute methods (MAMs) provide a wealth of information, and other techniques, such as HILIC and CE-MS, also continue to evolve.
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Cover Story - Technical Challenges Encountered During Chromatographic Method Transfers to Pharmaceutical Manufacturing SitesDrug substance development requires a range of analytical methods to be developed to generate process knowledge and to support in-process and release testing throughout a synthetic sequence. This article describes practical examples of a wide variety of transfer challenges and our remediation strategy.
