Affinity purification of recombinant interferon-α on a mimetic ligand adsorbent

被引：28

作者：

Swaminathan, S ^{[1
]}

Khanna, N ^{[1
]}

机构：

[1] Int Ctr Genet Engn & Biotechnol, Recombinant Gene Prod Lab, New Delhi 110067, India

来源：

PROTEIN EXPRESSION AND PURIFICATION | 1999年 / 15卷 / 02期

关键词：

interferon-alpha; purification; refolding; dye-affinity chromatography; downstream processing;

D O I：

10.1006/prep.1998.1017

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A method for improved refolding and purification of recombinant human interferon-alpha (rh-IFN-alpha) from inclusion bodies is described, The optimal conditions of refolding were obtained by the addition of 0.5 M L-argnine to the refolding buffer. The rh-IFN-alpha was purified to near homogeneity utilizing a single-step chromatography on a mimetic dye ligand matrix. Improved refolding, coupled to a single-column affinity purification strategy, resulted in a 10-fold increase in the yield of rh-IFN-alpha. This single-step purification protocol yielded similar to 50 mg of purified rh-IFN-alpha from 1 liter of shake flask culture. The rh-IFN-alpha prepared by this protocol was found to be essentially monomeric based on HPLC gel filtration and nonreducing SDS-PAGE, It had a specific activity of similar to 2.8 x 10(8) IU/mg, measured as inhibition of cytopathic effect of encephalomyocarditis virus on A549 human lung carcinoma cells. (C) 1999 Academic Press.

引用

页码：236 / 242

页数：7

共 20 条

[1] Method for increasing the yield of properly folded recombinant human gamma interferon from inclusion bodies [J].

Arora, D ;

Khanna, N .

JOURNAL OF BIOTECHNOLOGY, 1996, 52 (02) :127-133

[2]

BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3

[3] A METHOD FOR INCREASING THE YIELD OF PROPERLY FOLDED RECOMBINANT FUSION PROTEINS - SINGLE-CHAIN IMMUNOTOXINS FROM RENATURATION OF BACTERIAL INCLUSION-BODIES [J].

BUCHNER, J ;

PASTAN, I ;

BRINKMANN, U .

ANALYTICAL BIOCHEMISTRY, 1992, 205 (02) :263-270

[4]

CHO JM, 1993, Patent No. 0553494

[5]

Clark EDB, 1998, CURR OPIN BIOTECH, V9, P157

[6] CHEMICAL SYNTHESIS OF A HUMAN INTERFERON-ALPHA-2 GENE AND ITS EXPRESSION IN ESCHERICHIA-COLI [J].

EDGE, MD ;

GREENE, AR ;

HEATHCLIFFE, GR ;

MOORE, VE ;

FAULKNER, NJ ;

CAMBLE, R ;

PETTER, NN ;

TRUEMAN, P ;

SCHUCH, W ;

HENNAM, J ;

ATKINSON, TC ;

NEWTON, CR ;

MARKHAM, AF .

NUCLEIC ACIDS RESEARCH, 1983, 11 (18) :6419-6435

[7]

ETTLIN U, 1995, Patent No. 0679718

[8] CYTOKINE THERAPEUTICS - LESSONS FROM INTERFERON-ALPHA [J].

GUTTERMAN, JU .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1994, 91 (04) :1198-1205

[9] Strategies for optimizing heterologous protein expression in Escherichia coli [J].

Hannig, G ;

Makrides, SC .

TRENDS IN BIOTECHNOLOGY, 1998, 16 (02) :54-60

[10] EXPRESSION OF A HUMAN-GENE FOR INTERFERON IN YEAST [J].

HITZEMAN, RA ;

HAGIE, FE ;

LEVINE, HL ;

GOEDDEL, DV ;

AMMERER, G ;

HALL, BD .

NATURE, 1981, 293 (5835) :717-722

← 1 2 →