Webinar Review: Bioanalysis Of Therapeutic and Endogenous Peptides: Understanding and Overcoming the Challenges of Working with Large Molecules

Over the course of the semester, our trainees are reviewing webinars in their given fields and preparing abstracts to help colleagues outside their discipline make an informed choice about watching them. As our program bridges diverse disciplines, these abstracts are beneficial for our own group in helping one another gain key knowledge in each other’s fields. We are happy to share these here for anyone else who may find them helpful.

Bioanalysis Of Therapeutic and Endogenous Peptides: Understanding and Overcoming the Challenges of Working with Large Molecules

Erin Chambers


Waters Corporation webinars

Watch on the Waters website >>

Samuel OkyemAnalysis by Samuel Okyem: 

Endogenous peptides play a significant role in neuronal communication, making it a major target either as disease biomarkers or a therapeutical agent. Recent analysis estimates 60% of therapeutics believed to be biologics (e.g., peptides and proteins) in the next few decades. High throughput and efficient quantitative method are therefore needed in peptide measurement.

In this webinar, Erin Chambers, a principal application scientist at Waters Corporation, examines a step-by-step approach for effective quantitation of endogenous and therapeutic peptides.

Liquid chromatography coupled with mass spectrometry (LC-MS) is currently the most effective analytical tool in peptide measurement. Chromatographic conditions as well mass spectrometric approach needs to be optimized to enhance reproducibility and improve high peptide coverage.  Chambers discussed several chromatographic and mass spectrometry that led to improved reproducibility, coverage, and accuracy in quantitative peptidomics.

She reviewed how choosing a spectrometry with a high mass range allows the selection of precursor ions with lower charges which is proven to be more reproducible compared to the high charged state species. Also, she contended that fragment immonium ions may look very intense in a mass spectrum but should be avoided in quantitation since it is not reproducible and its unspecific. Chromatographic conditions also contributes significantly to the accuracy, reproducibility, and throughput of peptide measurement. Here, she elaborated on how column dimensions and particle properties can affect peak shape and resolution. Thus, column with smaller particle sizes and larger pore sizes improves resolution of peptides.

A major challenge in peptide analysis is matrix effect, which may result in low peptide recovery during solid phase extraction. An improved peptide recovery efficiency was observed when a generic C-18 column was replaced with a C-18 column modified with some charged molecules. In summary, these optimizations improved peptide measurement and was used to successfully quantify beta amyloids in biological fluids. The webinar gives a detailed insight on how you can improve your analytical performance in peptide detection and quantitation by selecting the proper mass spectrometry, chromatographic column and sample preparation method.