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Updated: Jan 28, 2026
06:26
Rapid Assessment of Membrane Protein Quality by Fluorescent Size Exclusion Chromatography
Published on: January 6, 2023
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Robust aggregate separation by Sartobind Rapid A Protein A membrane.
Gaoya Yuan1, Meng Qu1, Yifeng Li1
1Downstream Process Development, (DSPD), WuXi Biologics, 31 Yiwei Road, Waigaoqiao Free Trade Zone, Shanghai, 200131, China.
Protein Expression and Purification
|January 26, 2026
Summary
Protein A membrane effectively separates monomers from aggregates in antibody purification, even with high aggregate levels. This robust performance makes it superior to Protein A columns for challenging samples.
Area of Science:
- Biotechnology
- Bioprocessing
- Protein Purification
Background:
- Protein A resins are standard for antibody and Fc-fusion protein capture but struggle with monomer-aggregate separation.
- Protein A membranes offer potential advantages over traditional resin-based columns.
- Previous work showed Sartobind Rapid A membrane outperforms resin-based Protein A columns in aggregate separation.
Purpose of the Study:
- To evaluate mobile phase additives (NaCl, CaCl2, Arg·HCl) for optimizing monomer-aggregate separation using Sartobind Rapid A membrane.
- To assess the aggregate removal efficiency of Sartobind Rapid A membrane across various aggregate concentrations.
Main Methods:
- Tested three mobile phase additives to determine optimal conditions for monomer-aggregate separation.
- Challenged Sartobind Rapid A membrane with artificial samples containing 15-60% aggregates.
- Analyzed elution peaks using size-exclusion chromatography-high performance liquid chromatography (SEC-HPLC).
Main Results:
- Identified optimal mobile phase additive conditions for enhanced monomer-aggregate resolution.
- Sartobind Rapid A membrane effectively removed the majority of aggregates from all tested samples.
- High aggregate removal efficiency was consistent across samples with varying aggregate percentages (15-60%).
Conclusions:
- Sartobind Rapid A membrane demonstrates effective and robust aggregate separation capabilities.
- Protein A membrane chromatography is a superior alternative to Protein A columns for product capture, particularly with high-aggregate feed streams.
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