Continuous-bed chromatography for the analysis and purification of recombinant human basic fibroblast growth factor

被引：11

作者：

Garke, G ^{[1
]}

Radtschenko, I ^{[1
]}

Anspach, FB ^{[1
]}

机构：

[1] Gesell Biotechnol Forsch GmbH, Div Biochem Engn, D-38124 Braunschweig, Germany

来源：

JOURNAL OF CHROMATOGRAPHY A | 1999年 / 857卷 / 1-2期

关键词：

growth factors; continuous beds; proteins;

D O I：

10.1016/S0021-9673(99)00786-4

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

The chromatographic properties of the commercial cation exchanger UNO-Si (35x7 mm) was investigated using lysozyme from hen egg white as model protein and recombinant human basic fibroblast growth factor (rh-bFGF) from a high cell density cultivation of E. coli. The dynamic capacity for lysozyme (c degrees=1 mg/ml) in 100 mM acetate buffer, pH 5 was 27 mg per mi sorbent. It was found independent of the how-rate from 78 to 935 cm/h owing to the absence of mass transfer restrictions with this column concept. Regarding the selectivity for rh-bFGF and the capacity for lysozyme, no changes were apparent after cleaning-in-place (CIP) procedures with 0.5 M NaOH. Clogging of the column by a clarified crude cell homogenate of E. coli was not critical as precipitates were removed by reversal of the flow during CIP. Rh-bFGF elutes in three consequent peaks from the UNO-S1 column, which could be attributed to soluble rh-bFGF aggregates of different size. The dynamics of rh-bFGF aggregation and reaggregation in the crude feedstock was monitored by fast gradient elution chromatography. (C) 1999 Elsevier Science B.V. All rights reserved.

引用

页码：137 / 144

页数：8

共 22 条

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