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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
Author(s) -
Gerstmayer Bernhard,
Pessara Ulrich,
Wels Winfried
Publication year - 1997
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(97)00294-9
Subject(s) - pichia pastoris , recombinant dna , yeast , extracellular , pichia , biology , receptor , microbiology and biotechnology , biochemistry , chemistry , gene
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.