Construction and purification of domain-deleted immunoglobulin variants of the recombinant/chimeric B72.3 (y1) monoclonal antibody.
Chimeric antibodies have been produced against a pancarcinomic tumor associated antigen, TAG-72, by fusing the genes for the variable region of mouse MAb B72.3 to the genes for the constant region of human IgG. In our efforts to optimize the pharmacokinetics of plasma clearance and the efficiency of tumor localization and penetrance of cB72.3, we have now developed truncated versions of immunoglobulin heavy chains. The domain-deleted antibodies are produced by transfecting cells that produce chimeric kappa chains with expression vectors that encode chimeric heavy chains lacking the sequences that encode the CH2 domain, CH3 domain, or both. Despite the absence of these domains, the transfectomas secrete H2L2 tetramers with appropriate antigenic specificity. All the domain-deleted immunoglobulins can be purified by chromatography on Protein G Sepharose which binds to a site on the Fab region that is retained in the domain-deleted antibodies. The CH2CH3 domain-deleted immunoglobulin produced in cell culture is analogous in size to enzymatically produced F(ab')2.
Calvo
B
B
Laboratory of Tumor Immunology and Biology, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892.
Kashmiri
S V
SV
Hutzell
P
P
Hand
P H
PH
Slavin-Chiorini
D C
DC
Schlom
J
J
Zaremba
S
S
D000906:Antibodies / Q000378:metabolism; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000235:genetics* / Q000302:isolation & purification; D000951:Antigens, Neoplasm / Q000276:immunology; D001483:Base Sequence; D001666:Binding Sites, Antibody; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes; D005803:Genes, Immunoglobulin; D006023:Glycoproteins / Q000276:immunology; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D007147:Immunoglobulin Light Chains / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D007136:Immunoglobulins / Q000096:biosynthesis* / Q000235:genetics* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics* / Q000302:isolation & purification; D014162:Transfection
D000906:Antibodies; D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D000939:Epitopes; C029152:G-substrate; D006023:Glycoproteins; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007136:Immunoglobulins; D009419:Nerve Tissue Proteins; D011993:Recombinant Fusion Proteins; C047052:tumor-associated antigen 72