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.
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.
Gerstmayer
B
B
Institute for Experimental Cancer Research, Tumor Biology Center, Freiburg, Germany.
Pessara
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U
Wels
W
W
D000069594:Abatacept; D000818:Animals; D015703:Antigens, CD / Q000096:biosynthesis* / Q000235:genetics; D000943:Antigens, Differentiation / Q000096:biosynthesis* / Q000235:genetics; D018122:B7-1 Antigen / Q000096:biosynthesis* / Q000235:genetics; D051940:B7-2 Antigen; D001681:Biological Assay; D060908:CTLA-4 Antigen; D019281:Dimerization; D006801:Humans; D018796:Immunoconjugates; D007166:Immunosuppressive Agents; D008213:Lymphocyte Activation; D008562:Membrane Glycoproteins / Q000096:biosynthesis* / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D010446:Peptide Fragments / Q000096:biosynthesis; D010843:Pichia / Q000235:genetics; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000096:biosynthesis
D015703:Antigens, CD; D000943:Antigens, Differentiation; D018122:B7-1 Antigen; D051940:B7-2 Antigen; C497964:CD86 protein, human; D060908:CTLA-4 Antigen; C556706:CTLA4 protein, human; C497965:Cd86 protein, mouse; C556707:Ctla4 protein, mouse; D018796:Immunoconjugates; D007166:Immunosuppressive Agents; D008562:Membrane Glycoproteins; D010446:Peptide Fragments; D011994:Recombinant Proteins; D000069594:Abatacept