Baculovirus Recombinant Expressing a Secreted Form of a Transmembrane Carcinoma-associated Antigen1

GA733-2 is a monoclonal antibody-defined, 40-kDa glycoprotein an tigen that is associated with carcinomas of various origins. Hydrophobicity analysis of the protein sequence predicted by complementary DNA (cDNA) has suggested that the GA733-2 antigen is a type I membrane protein. In this study, the polymerase chain reaction was used in a strategy to omit cDNA sequences for the transmembrane and cytoplasmic domains, thereby converting the extracellular domain into a secretory protein. Full-length and truncated cDNAs were cloned into the baculovirus transfer vector pVL1392 and introduced into Autographa califor nien nuclear polyhedrosis virus by homologous recombination. The fulllength cDNA baculovirus recombinant directed the expression of a 40kDa glycoprotein that was confined to infected Spodoptera frugiperda cells, whereas cells infected with the truncated cDNA baculovirus recom binant abundantly secreted a 31-kDa glycoprotein into the culture me dium. Recombinant secretory antigen displayed an in vitro immunoreactivity to monoclonal antibody and an in vivo immunogenicity in mice that were similar to native antigen. The facile purification of mg quantities of carcinoma-associated antigen will enable an evaluation of its immunoge nicity in cancer patients.