Cell‐free immunology: construction and in vitro expression of a PCR‐based library encoding a single‐chain antibody repertoire

FEBS Letters

Spirin, Alexander S.¹  Kolb, Vyacheslav A.¹  Makeyev, Eugeny V.¹ 

[1] Institute of Protein Research, Russian Academy of Sciences, 142292 Pushchino, Moscow Region, Russia

关键词: Single-chain antibody;    Polymerase chain reaction library;    Cell-free translation;    Ribosome;    BCIP;    5-bromo-4-chloro-3-indolyl-phosphate;    K;    kappa light chain;    NBT;    4-nitroblue tetrazolium chloride;    PCR;    polymerase chain reaction;    scAb;    single-chain antibody;    SDS;    sodium dodecyl sulfate;    SOE;    splicing by overlap extension;    VH;    heavy chain variable domain;   

DOI  :  10.1016/S0014-5793(99)00055-1

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

A novel cloning-independent strategy has been developed to generate a combinatorial library of PCR fragments encoding a murine single-chain antibody repertoire and express it directly in a cell-free system. The new approach provides an effective alternative to the techniques involving in vivo procedures of preparation and handling large libraries of antibodies. The possible use of the described strategy in the ribosome display is discussed.

【 授权许可】

Unknown