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A Lectin HPLC Method to Enrich Selectively-glycosylated Peptides from Complex Biological Samples
Published on: October 1, 2009
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Multi-arm PEG-maleimide conjugation intermediate characterization and hydrolysis study by a selective HPLC method.
Jenny Wang1, Samuel Yang1, Kelly Zhang1
1Research and Early Development, Genentech Inc., 1 DNA Way, South San Francisco, CA 94080, United States.
Journal of Pharmaceutical and Biomedical Analysis
|November 18, 2018
Summary
A new HPLC method effectively characterizes 8-arm polyethylene glycol (PEG)-maleimide, crucial for drug conjugation. This stability-indicating method analyzes hydrolysis, guiding optimized protein conjugation processes.
Area of Science:
- Polymer Chemistry
- Analytical Chemistry
- Bioconjugation Chemistry
Background:
- Maleimide-functionalized polyethylene glycol (PEG) is vital for drug conjugation.
- Characterizing high molecular weight PEG-maleimide and its hydrolysis products in solution is analytically challenging.
- Understanding PEG-maleimide stability is critical for successful drug delivery applications.
Purpose of the Study:
- To develop a highly selective, sensitive, and stability-indicating HPLC method for characterizing 8-arm PEG-maleimide.
- To investigate the hydrolysis of 8-arm PEG-maleimide under various conditions.
- To determine the kinetics of PEG-maleimide hydrolysis to inform process development.
Main Methods:
- Development and validation of a High-Performance Liquid Chromatography (HPLC) method using a charged surface hybrid (CSH) stationary phase.
- Method validation according to ICH Q2 guidelines for specificity, sensitivity, linearity, accuracy, and precision.
- Study of 8-arm PEG-maleimide hydrolysis in different buffer media, pH, and temperatures.
Main Results:
- A robust HPLC method was established for 8-arm PEG-maleimide characterization and quality control.
- Hydrolysis of 8-arm PEG-maleimide was confirmed as a first-order reaction.
- Kinetic parameters, including the hydrolysis rate constant and half-life, were determined.
Conclusions:
- The developed HPLC method provides reliable characterization of 8-arm PEG-maleimide, essential for quality and process control.
- Understanding PEG-maleimide hydrolysis kinetics is crucial for optimizing protein conjugation processes.
- This research supports the development of stable PEGylated biotherapeutics.
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