Effect of cross-reactivity of alpha-fetoprotein monoclonal antibody on quantitation of serum AFP and radioimmunodetection of hepatocellular carcinoma.
Murine hybridomas were generated by immunizing mice with purified alpha-fetoprotein (AFP) to determine the cross-reactivity of AFP antibodies with human serum albumin (HSA). Eighteen out of 23 hybridoma products were cross-reactive with HSA. All 23 monoclonal antibodies (MAbs) could be subgrouped into 3 groups by their binding affinity to AFP and HSA, and one MAb from each group, IIB3, IIIB1 and IIIC6, were chosen for this study. The affinity to AFP was highest for IIB3 followed by IIIC6 and IIIB1, and that to HSA was in the order IIIB1 greater than IIB3. Immunohistochemical staining revealed no cross-reactions of these antibodies with HSA at the tissue level. IIB3 was the most efficient in the quantitative assays, indicating high affinity and specificity (non-cross-reactivity) of the antibodies, both of which are prerequisites for in vitro assays. Scans with good tumor localization were obtained from mice which received 131I-labeled IIB3 or IIIC6 after xenografting of the human hepatocellular carcinoma. The images obtained with IIB3 were inhibited in the presence of either AFP or HSA in the circulation. These findings indicated that the use of cross-reactive and/or high-affinity antibodies against AFP provides an inhibitory factor for tumor localization. MAb IIIC6, because of its high specificity and moderate affinity to AFP, seems to be the best candidate for immunoscintigraphy of the tumors in future studies.
Makidono
R
R
Department of Radiology, Faculty of Medicine, Kyushu University, Fukuoka, Japan.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000915:Antibody Affinity; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D006825:Hybridomas; D007150:Immunohistochemistry; D008114:Liver Neoplasms, Experimental / Q000032:analysis / Q000000981:diagnostic imaging*; D051379:Mice; D008807:Mice, Inbred BALB C; D008970:Molecular Weight; D011233:Precipitin Tests; D011863:Radioimmunoassay; D011877:Radionuclide Imaging; D012709:Serum Albumin / Q000276:immunology*; D014407:Tumor Cells, Cultured; D000509:alpha-Fetoproteins / Q000032:analysis / Q000276:immunology*
D000911:Antibodies, Monoclonal; D012709:Serum Albumin; D000509:alpha-Fetoproteins