Peptide and Protein Bioanalysis Boot Camp

Building Proficiency in LC-MS Large Molecule Quantification

Peptide and Protein Bioanalysis Boot Camp

Waters’ team of bioanalysis experts developed the Peptide & Protein Bioanalysis Boot Camp – a series of short, recorded presentations – to help you and your colleagues acquire a fundamental and practical proficiency in large molecule quantification.

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Introduction to Peptide & Protein Bioanalysis Boot Camp

Quantifying large molecules in matrix is new to many of us and presents a variety of challenges that we don’t see with small molecules. Peptides and proteins have a more diverse set of chemical properties and tend to be much more complex. Kelly Doering, Senior Business Manager, Bioanalysis & DMPK, explains the goals of the Peptide and Protein Bioanalysis Bootcamp.
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Introduction to Peptides and Proteins for Bioanalysis Using LC-MS

Khalid Khan, Senior Manager Business Development, discusses the basic structure of amino acids, peptides, and proteins, including specific examples like monoclonal antibodies. The ionization and fragmentation of peptides and how they differ from small molecules is also discussed.
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Contrasting LBAs and LC-MS for Peptide and Protein Bioanalysis

Kelly Doering discusses ligand binding assays (LBAs) and compares the advantages and limitations of this technology to LC-MS. She'll then present three examples of when LC-MS is the preferred method for quantification of insulin analogs based on specificity requirements and the need to multiplex.
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LC-MS/MS for Bioanalytical Peptide and Protein Quantification: MS Considerations

Caitlin Dunning, Associate Scientist, discusses how to use mass spectrometry to develop sensitive, selective, and robust quantification methods for peptide and protein quantification, including: basic mass spectrometry considerations, MS tuning, and best practices for success. In addition, practical considerations are provided to achieve greater MRM specificity for your peptides of interest.
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LC-MS/MS for Bioanalytical Peptide and Protein Quantification: Chromatographic Considerations

Bioanalytical scientists are faced with developing robust, reliable, and sensitive methods. This is especially challenging when we deal with peptides and proteins. Mary Lame, Principal Applications Chemist, walks you through a basic LC screening method designed specifically with peptides in mind.
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LC-MS/MS for Bioanalytical Peptide and Protein Quantification: Peptide Level Sample Clean-up

Bioanalytical laboratories are continually pressed to attain increasing levels of performance and productivity in the development of robust, reliable, and sensitive methods for accurate quantification. The challenge is the high sensitivity quantification of peptides and proteins from complex biological matrices. Mary Lame, Principal Applications Chemist, presents the starting universal solid phase extraction protocol for therapeutic, endogenous, and tryptic peptides. The value of incorporating a mixed mode SPE strategy into the overall LC-MS quantitative workflow is demonstrated.
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Peptide Sample Preparation Optimization and Troubleshooting

Mass spectrometry, chromatography, and sample preparation are all leveraged to facilitate the development of sensitive, selective, and robust bioanalytical methods for the quantification of peptides in biological matrices. In this module, Erin Chambers, Vice President, Chemistry and Technology, focuses on peptide sample prep optimization and the fundamental characteristics of peptides.
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Identification of Surrogate Peptides for LC-MS Protein Quantification

In this module Nikunj Tanna, Waters' Senior Scientist, discusses the circumstances in which it's necessary to perform a surrogate peptide approach for protein quantification; the differences between generic and unique surrogate peptides; and he outlines various steps in identifying peptides, and the available software tools to aid the process.
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MRM Optimization for Surrogate Peptides for LC-MS Protein Quantification

Paula Orens, Application Scientist, examines a typical workflow for surrogate peptide approach via LC-MS with tandem quadrupole mass spectrometry. Identifying transitions and optimizing MS conditions is a key focus of this module. By the end, you will understand how mass spectrometry informatics systesm combined with an open-source client MRM application can work together to simplify MRM method development. Using Infliximab as an example, Ms. Orens demonstrates the development and optimization of MRM transitions to establish sensitivity needs and discusses sample prep and LC considerations.