Single-chain Fv with Fc fragment of the human IgG1 tag:: Construction, Pichia pastoris expression and antigen binding characterization

被引：23

作者：

de Andrade, EV ^{[1
]}

de Albuquerque, FC ^{[1
]}

de Moraes, LMP ^{[1
]}

Brígido, MD ^{[1
]}

Santos-Silva, MA ^{[1
]}

机构：

[1] Univ Brasilia, Mol Biol Lab, Brasilia, DF, Brazil

来源：

JOURNAL OF BIOCHEMISTRY | 2000年 / 128卷 / 06期

关键词：

antibody engineering; antibody fragment; antigen binding; Pichia pastoris; single-chain Fv;

D O I：

10.1093/oxfordjournals.jbchem.a022838

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

We report two expression vectors in Pichia pastoris that direct the synthesis of recombinant single chain antibody variable region (scFv), derived from anti-Z-DNA monoclonal antibody Z22, The first vector codes for a scFv fused to the Ig binding domain of staphylococcal Protein A. The second vector codes for the scFv fused to the Fc fragment of the human IgG1, The fusion partner simplified the detection and purification of the secreted protein. These constructs yielded high level expression of an scFv with specific binding activity toward a Z form of DNA, with binding activity comparable to that of the scFv molecule produced in an Escherichia coli expression system and the original monoclonal antibody.

引用

页码：891 / 895

页数：5

共 13 条

[1]

BRIGIDO MM, 1993, J IMMUNOL, V150, P469

[2] YEAST SYSTEMS FOR THE COMMERCIAL PRODUCTION OF HETEROLOGOUS PROTEINS [J].