A simple method for midkine purification by affinity chromatography with a heavy chain variable domain (VH) fragment of antibody
A simple method for midkine purification by affinity chromatography with a heavy chain variable domain (VH) fragment of antibody
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DOI：
10.1016/s0165-022x(00)00145-7
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发表时间：
2001-03-28
影响因子：
--
通讯作者：
Shinozawa, T
中科院分区：
文献类型：
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作者：
Dansithong, W;Paul, S;Shinozawa, T
从产生抗人中期因子（MK）的单克隆抗体的杂交瘤制备重链可变结构域（VH）的DNA片段。在大肠杆菌中产生抗体片段，并通过ELISA证实其对化学合成的全长MK或重组中期因子C-末端（MKC-半）蛋白的亲和力。将表达MKc-半的大肠杆菌细胞裂解物施加到VH片段偶联的Sepharose 4 B柱上，并用含有0.5M NaCl的缓冲液洗脱。SDS-PAGE分析显示洗脱液中MKc半蛋白的纯度很高，表明重组VH片段在通过亲和层析纯化蛋白质中的实用性。(C)2001 Elsevier Science B. V.保留所有权利。
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.