A simple method for midkine purification by affinity chromatography with a heavy chain variable domain (VH) fragment of antibody

被引：6

作者：

Dansithong, W ^{[1
]}

Paul, S ^{[1
]}

Kojima, Y ^{[1
]}

Kamiya, K ^{[1
]}

Shinozawa, T ^{[1
]}

机构：

[1] Gunma Univ, Fac Engn, Dept Biol & Chem Engn, Kiryu, Gumma 3768515, Japan

来源：

JOURNAL OF BIOCHEMICAL AND BIOPHYSICAL METHODS | 2001年 / 48卷 / 01期

关键词：

affinity chromatography; midkine (MK); protein purification; variable heavy chain (VH);

D O I：

10.1016/S0165-022X(00)00145-7

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A DNA fragment for a heavy chain variable domain (VH) was prepared from a hybridoma that produces a monoclonal antibody against human midkine (MK). The antibody fragment was produced in Escherichia coli and its affinity for chemically synthesized full length MK or recombinant midkine c-terminus (MKc-half) protein was confirmed by ELISA. An Escherichia coli cell lysate expressing MKc-half was applied to a VH fragment-coupled Sepharose 4B column and eluted with a buffer containing 0.5 M NaCl. SDS-PAGE analysis revealed a high degree of purity of the MKc-half protein in the eluent, showing the utility of a recombinant VH fragment in purification of proteins by affinity chromatography. (C) 2001 Elsevier Science B.V. All rights reserved.

引用

页码：77 / 84

页数：8

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