Patent ID: 8067197

Claim:
A process for preparing a large amount of a purified soluble recombinant human α V β 3 adhesion receptor comprising an α V chain and a β 3 chain and having substantially unimpaired ligand binding activity, comprising (i) subcloning (a) a first cDNA coding for the α V chain of said receptor, truncated by a portion comprising at least 61 amino acids from the C-terminus of a mature human α V protein comprising approximately 1018 amino acids, and (b) a second cDNA coding for the β 3 chain of said receptor, truncated by a portion comprising at least 70 amino acids from the C-terminus of a mature human β 3 protein chain comprising approximately 762 amino acids, into a baculovirus transfer vector of a baculovirus expression system, (ii) transferring said vector comprising said first and/or second DNA into the genomic DNA of a baculovirus of said expression system, (iii) infecting an insect cell with said complete recombinant baculovirus, (iv) cultivating said infected insect cells in a culture medium whereby said truncated α V β 3 receptor is expressed into the medium, and (v) purifying said expressed receptor from the medium by antibody affinity chromatography, wherein the antibody is specific to the human α V β 3 adhesion receptor or its individual component chains, wherein the cDNA of the truncated α V chain is generated by PCR using the oligonucleotide primers 5′-GAC CAG CAT TTA CAG TGA-3′ (SEQ ID NO: 4) and 5′-CA CAG GTC TAG ACT ATG GCT GAA TGC CCC AGG-3′ (SEQ ID NO: 5), and the cDNA of the truncated β 3 chain is generated by PCR using the oligonucleotide primers 5′-GCG CGC AAG CTT GCC GCC ACC ATG CGA GCG CGG CCG-3′ (SEQ ID NO: 6) and 5′GAT CGA TCT AGA CTA GGT CAG GGC CCT TGG GAC ACT-3′ (SEQ ID NO: 7).