Protein charge homogeneity can have a significant effect on the structure, stability, binding affinity, and efficacy of a biotherapeutic drug. Speed and accuracy are key criteria for laboratories performing charge variant profiling. Especially in contract research organizations who deliver confident, fast analytical services to the biopharmaceutical and biosimilar industry. Ion exchange chromatography (IEX) is often used for protein charge variant profiling, either by salt gradient or pH gradient and can be easily implemented for routine analysis with buffer kits, correct column selection and easy-to-use software.


Charge variant analysis workflow

Identification and separation of charge variants is performed rapidly and reliably with full Vanquish UHPLC solutions. Separations of monoclonal antibody variants can be challenging as they are complex molecules and can differ by as little as one charge. The use of a pH gradient-based elution can be easily implemented with buffer kits and lead to  faster, more robust and less challenging separations than salt-based elutions with better resolution and sensitivity.  Additionally the column chemistry is critical for optimized separations. The unique nonporous pellicular resin used in MAbPac SCX-10 provides high resolving power, permitting the separation of monoclonal antibody variants that differ by as little as one charge.


Resources

Type Title
Application Note
Application Note  High Salt Gradient Analysis of Post-Translational Modifications - Deamidation Monitoring
Application Note  Utilizing the Native Fluorescence of Monoclonal Antibodies for the Sensitive Detection of Charge Variants
Application Note  Transferring a Method for the Characterization of PTMs from the UltiMate 3000 BioRS System to the Vanquish Flex UHPLC System
Application Note  Development of Ultra-fast pH-Gradient Ion Exchange Chromatography for the Separation of Monoclonal Antibody Charge Variants
Application Note  The advantage of pH gradient buffers is demonstrated by the ion exchange separation of charge variants of denosumab
Application Note  High-Resolution Charge Variant Analysis for Top-Selling Monoclonal Antibody Therapeutics Using a Linear pH Gradient Separation Platform
Application Note  Simultaneous analysis of monoclonal antibodies using a novel dual-channel UHPLC instrument and orthogonal chromatography
Application Note  Doubling the throughput of long chromatographic methods by using a novel Dual LC workflow
Charge Variant Methods  AppsLab Library - Charge Variant Methods
Customer Case Study  Bringing Biosimilar Therapeutics to Market Faster – A CRO Case Study
Customer Case Study
Webinar Series  Get Greater Structural Insight into Biotherapeutic Proteins Charge Variants and Aggregates
Video

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