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J Sep Sci. 2017 Apr;40(7):1493-1499. doi: 10.1002/jssc.201601196. Epub 2017 Mar 15.

Enhanced binding by dextran-grafting to Protein A affinity chromatographic media.

Author information

1
State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing, P.R. China.
2
School of Chemical and Environmental Engineering, China University of Mining and Technology, Beijing, P.R. China.
3
Department of Chemical Engineering, Beijing Institute of Petro-chemical Technology, Beijing, P.R. China.
4
Jiangsu National Synergetic Innovation, Center for Advance Materials (SICAM), Nanjing, P.R. China.

Abstract

Dextran-grafted Protein A affinity chromatographic medium was prepared by grafting dextran to agarose-based matrix, followed by epoxy-activation and Protein A coupling site-directed to sulfhydryl groups of cysteine molecules. An enhancement of both the binding performance and the stability was achieved for this dextran-grafted Protein A chromatographic medium. Its dynamic binding capacity was 61 mg immunoglobulin G/mL suction-dried gel, increased by 24% compared with that of the non-grafted medium. The binding capacity of dextran-grafted medium decreased about 7% after 40 cleaning-in-place cycles, much lower than that of the non-grafted medium as decreased about 15%. Confocal laser scanning microscopy results showed that immunoglobulin G was bound to both the outside and the inside of dextran-grafted medium faster than that of non-grafted one. Atomic force microscopy showed that this dextran-grafted Protein A medium had much rougher surface with a vertical coordinate range of ±80 nm, while that of non-grafted one was ±10 nm. Grafted dextran provided a more stereo surface morphology and immunoglobulin G molecules were more easily to be bound. This high-performance dextran-grafted Protein A affinity chromatographic medium has promising applications in large-scale antibody purification.

KEYWORDS:

Protein A; affinity chromatography; antibody purification; dextran; grafting

PMID:
28234424
DOI:
10.1002/jssc.201601196
[Indexed for MEDLINE]

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