Selection and characterisation of binders based on homodimerisation of immunoglobulin VH domains

FEBS Letters

Jin, Hulin¹  Burrone, Oscar R.¹  Sepúlveda, Jorge¹ 

[1] International Centre for Genetic Engineering and Biotechnology, Area Science Park, Padriciano 99, 34012 Trieste, Italy

关键词: Antibody engineering;    Heavy chain variable domain;    Phage display;    pIII;   

DOI  :  10.1016/S0014-5793(03)01182-7

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The antigen-binding surface of antibodies is formed by the heterodimerisation of the two variable domains of the light (V_L) and heavy (V_H) chains. We have previously described the spontaneous formation of V_H dimers (VHD) in both bacteria and mammalian cells. The self-association of a single domain produces a homo-VHD, in which the two identical V_H domains generate a unique symmetric surface for antigen binding that is never found in the normal V_L/V_H antibody binding site. We developed a phagemid vector for the construction of phage display libraries in which a cysteine residue, introduced at the C-terminus of the only V_H cloned, allowed display of homo-VHDs. Panning of the library on different proteins yielded antigen specific binders against lysozyme, glutathione S-transferase and streptavidin. A lysozyme specific homo-VHD was further characterised with an apparent affinity determined to be 216±6.6 nM. Importantly, the results showed that its binding activity was fully dependent on the dimerisation of both identical V_H domains.

【 授权许可】

Unknown