Construction and expression in the yeast Pichia pastoris of functionally active soluble forms of the human costimulatory molecules B7‐1 and B7‐2 and the B7 counter‐receptor CTLA‐4

FEBS Letters

Pessara, Ulrich¹  Wels, Winfried²  Gerstmayer, Bernhard² 

[1] Boehringer Mannheim GmbH, Nonnenwald 2, D-82372 Penzberg, Germany;Institute for Experimental Cancer Research, Tumor Biology Center, Breisacher Straße 117, D-79106 Freiburg, Germany

关键词: Pichia pastoris;    Heterologous expression;    Human B7;    Human CTLA-4;    T cell costimulation;   

DOI  :  10.1016/S0014-5793(97)00294-9

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

We have generated soluble recombinant forms of the costimulatory molecules B7-1 and B7-2, and their counter-receptor CTLA-4 using a yeast Pichia pastoris expression system. Fragments comprising the extracellular domains of human B7-1, B7-2, and CTLA-4 molecules were expressed at high levels and could be purified from culture supernatants following a simple one-step purification protocol. The recombinant proteins retained their functionality and specific binding to their natural counterparts could be demonstrated by FACS analysis. In T cell proliferation assays costimulatory activity of immobilized B7-1 and B7-2 proteins in the presence of an anti-CD3 antibody was observed with the B7-1 protein being more potent than B7-2.

【 授权许可】

Unknown