Real-time control of antibody loading during protein A affinity chromatography using an on-line assay

被引：32

作者：

Fahrner, RL ^{[1
]}

Blank, GS ^{[1
]}

机构：

[1] Genentech Inc, Dept Recovery Sci, San Francisco, CA 94080 USA

来源：

JOURNAL OF CHROMATOGRAPHY A | 1999年 / 849卷 / 01期

关键词：

process control; preparative chromatography; immobilized proteins; antibodies; proteins;

D O I：

10.1016/S0021-9673(99)00539-7

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

We show that an on-line chromatographic assay can reliably control antibody loading in real-time during protein A affinity chromatography purification of a recombinant antibody from clarified Chinese hamster ovary cell culture fluid. The on-line assay directly sampled preparative column effluent and provided real-time measurement of antibody breakthrough during loading. The on-line assay used protein A immobilized on perfusion chromatography media, equilibrated with phosphate buffered saline at pH 7.2 and eluted with phosphate-buffered saline at pH 2.2. The assay reliably ended loading at 1% breakthrough with minimal yield loss. Reproducible yield and purity were obtained over 23 consecutive cycles. Yield remained constant while breakthrough capacity and the antibody concentration in the load changed. (C) 1999 Elsevier Science B.V. All rights reserved.

引用

页码：191 / 196

页数：6

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