RECOMBINANT HUMAN TRANSFORMING GROWTH-FACTOR-BETA - EXPRESSION BY CHINESE-HAMSTER OVARY CELLS, ISOLATION, AND CHARACTERIZATION

被引：19

作者：

BOURDREL, L ^{[1
]}

LIN, CH ^{[1
]}

LAUREN, SL ^{[1
]}

ELMORE, RH ^{[1
]}

SUGARMAN, BJ ^{[1
]}

HU, S ^{[1
]}

WESTCOTT, KR ^{[1
]}

机构：

[1] AMGEN INC, AMGEN CTR 2-1-A-223, THOUSAND OAKS, CA 91320 USA

来源：

PROTEIN EXPRESSION AND PURIFICATION | 1993年 / 4卷 / 02期

关键词：

D O I：

10.1006/prep.1993.1019

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A partial cDNA clone encoding the gene for human transforming growth factor-β1 (TGF-β1) was isolated from a human bladder carcinoma cell line (5637) cDNA library. Following restriction enzyme processing and ligation of synthetic oligonucleotide linkers, the gene was inserted into a plasmid and transfected into Chinese hamster ovary cells. Clonal selection and growth conditions resulted in a method for production of recombinant human TGF-β1 at 7 mg/liter in conditioned cell medium. Through a combination of low pH treatment, cation-exchange chromatography, and salt precipitation, the recombinant human TGF-β1 was purified in milligram amounts to =95% purity in a yield of about 36%. Purification to homogeneity was accomplished by chromatography on C18 silica gel. Amino acid analysis, N-terminal sequencing, and growth inhibition assays indicate identity with the molecule from human platelets. © 1993 Academic Press. All rights reserved.

引用

页码：130 / 140

页数：11

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