Overview
Charge variant analysis using ion-exchange chromatography for the detection of protein therapeutic modifications.Thorough characterization of monoclonal antibodies (mAbs) throughout the drug development and production processes allows for the detection of potential modifications that can affect the product’s efficacy, safety, and stability. Charge variant analysis using ion-exchange chromatography (IEC) separates proteins based on differences in charge, making it an ideal method for detecting these modified species. IEC is based on a stationary phase with a charged ligand on the surface that interacts with analytes of the opposite charge. The technique is divided between anion-exchange and cation-exchange chromatography. In anion-exchange, positively charged surface ligands interact with negatively charged analytes; in cation-exchange, negatively charged surface ligands interact with positively charged analytes. |
Recommended HIC HPLC columns for charge variant analysis
| ProPac Elite WCX10 HPLC columns | MAbPac SCX-10 HPLC columns | ProPac SAX-10 HPLC columns |
|---|---|---|---|
✓ Use this weak cation-exchange column for proteins with a pI > 6 ✓ Provides ultra-high resolution, speed, and efficiency ✓ Able to use pH or salt gradients | ✓ Provides complementary selectivity to weak cation-exchange columns ✓ Able to run faster gradients over a wider pH range Note 1: Separation profiles may be different between the weak and strong cation-exchange columns. Evaluate both to determine which offers the highest resolution for your charge variants. Note 2: To quickly develop a method on a LC-UV platform for proteins with a pI from 6-10, use this column in combination with linear Thermo Scientific CX-1 pH gradient buffers. | ✓ Use this strong anion-exchange column for proteins with a pI < 6 ✓ Provides ultra-high resolution, speed, and efficiency ✓ Able to use pH or salt gradients | |
HPLC column selection guide
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Resources
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Taking Charged Variant Analysis of Therapeutic Proteins to the Next Level
Understanding the Versatility and Challenges of Charge Variant Analysis for Biotherapeutics
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