Baculovirus recombinants expressing the human carcinoembryonic antigen gene.
Carcinoembryonic antigen (CEA), one of the most extensively studied human tumor-associated antigens, represents a potential target for passive as well as active immunotherapy. We describe here the first baculovirus recombinants expressing the human CEA gene. Eight baculovirus clones were isolated which expressed products of varying molecular weights; one clone, termed BVCEA-140, was shown to contain multiple CEA epitopes by reactivity to a panel of anti-CEA monoclonal antibodies. When purified protein isolated from this clone was deglycosylated, immunoreactive species ranging from M(r) 50,000 to M(r) 110,000 were found. Results of Southern blot analysis carried out on BVCEA-140 DNA were consistent with the hypothesis that these products result from the stable expression of variants which have recombined within the repeated domains of CEA. Other baculovirus recombinants expressing products comprising different portions of the CEA gene were also derived. One, termed BVCEA-35, was shown to be a recombination between the first 87 bases of domains I and III of the CEA gene. A variant, termed BVCEA-16, contained only the NH2-terminal domain of the CEA gene. Moreover, a recombinant expressing the closely related molecule nonspecific cross-reactive antigen was also derived. As shown here, commercially available preparations of CEA, which are derived from tumor biopsies or cell line supernatants, may contain nonspecific cross-reacting antigens and other contaminants. Thus, the recombinant CEA molecules described should have numerous uses including validation of the use of monoclonal antibodies as standards in CEA serum assays, the characterization of immune responses to CEA, the use as immunogen, and the study of structure function relationships.
Salgaller
M L
ML
Laboratory of Tumor Immunology and Biology, National Cancer Institute, NIH, Bethesda, Maryland 20892.
Bei
R
R
Schlom
J
J
Poole
D J
DJ
Robbins
P F
PF
D000818:Animals; D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm / Q000235:genetics* / Q000276:immunology; D016367:Baculoviridae; D001483:Base Sequence; D015153:Blotting, Western; D002272:Carcinoembryonic Antigen / Q000235:genetics* / Q000276:immunology; D015815:Cell Adhesion Molecules; D002478:Cells, Cultured; D003001:Cloning, Molecular; D006023:Glycoproteins / Q000235:genetics; D066298:In Vitro Techniques; D008562:Membrane Glycoproteins / Q000276:immunology*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009036:Moths; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D011994:Recombinant Proteins
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D002272:Carcinoembryonic Antigen; D015815:Cell Adhesion Molecules; D006023:Glycoproteins; D008562:Membrane Glycoproteins; D009838:Oligodeoxyribonucleotides; D011994:Recombinant Proteins