Optimization of elution salt concentration in stepwise elution of protein chromatography using linear gradient elution data. Reducing residual protein A by cation-exchange chromatography in monoclonal antibody purification

Journal of Chromatography. a
Takashi IshiharaShuichi Yamamoto

Abstract

Our simple method for optimization of the elution salt concentration in stepwise elution was applied to the actual protein separation system, which involves several difficulties such as detection of the target. As a model separation system, reducing residual protein A by cation-exchange chromatography in human monoclonal antibody (hMab) purification was chosen. We carried out linear gradient elution experiments and obtained the data for the peak salt concentration of hMab and residual protein A, respectively. An enzyme-linked immunosorbent assay was applied to the measurement of the residual protein A. From these data, we calculated the distribution coefficient of the hMab and the residual protein A as a function of salt concentration. The optimal salt concentration of stepwise elution to reduce the residual protein A from the hMab was determined based on the relationship between the distribution coefficient and the salt concentration. Using the optimized condition, we successfully performed the separation, resulting in high recovery of hMab and the elimination of residual protein A.

Citations

May 19, 2009·Journal of Chromatography. a·Katarzyna WrzosekMilan Polakovic
Feb 5, 2016·Journal of Molecular Recognition : JMR·Chih-Pei LinMilton T W Hearn
May 21, 2019·Preparative Biochemistry & Biotechnology·Lalita K Shekhawat, Anurag S Rathore

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