[A monoclonal antibody directed against an enediyne antitumor antibiotic and its preliminary application].
C1027, composed of an enediyne chromophore and an apoprotein of 110 amino acid residues, is a new highly potent macromolecular antitumor antibiotic. In order to prepare monoclonal antibodies (McAb) against C1027 by hybridoma technique, natural C1027 was inactivated by ultraviolet and coupled with human serum albumin, then immunized BALB/c mice. After cell fusion and screening by ELISA, a hybridoma secreting anti-C1027 McAb designated as F9 was obtained. McAb F9 specifically reacted with C1027 as determined by immunoblot assay. No obvious difference was observed between the reactivity of McAb F9 to natural C1027 and that of McAb F9 to ultraviolet inactivated C1027. This result indicates that the ultraviolet-sensitive chromophore of C1027 may not participate in the epitope for McAb F9. The isotype of F9 is IgG1 and its affinity constant was found to be (2.2 +/- 0.47) x 10(7) L.mol-1 according to Beatty's method. By clonogenic assay, McAb F9 neither affected the cytotoxicity of C1027 to KB cells nor blocked the activity of the chromophore. McAb F9 also specifically reacted with the recombinant truncated C1027 apoprotein in which 16 amino acid residues at the C terminus were deleted. This study suggests that F9 is a valuable McAb for the research of C1027 apoprotein engineering, C1027 related immunoconjugates and screening of new macromolecular antitumor substances with homology to the protein part of C1027.
Zhou
C S
CS
Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Uion Medical College, Beijing 100050.
Xu
L N
LN
Jiang
M
M
Zhen
Y S
YS
D000617:Aminoglycosides; D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D000903:Antibiotics, Antineoplastic / Q000494:pharmacology*; D000911:Antibodies, Monoclonal; D053281:Enediynes; D051379:Mice; D008807:Mice, Inbred BALB C; D014410:Tumor Stem Cell Assay / Q000379:methods
D000617:Aminoglycosides; D000900:Anti-Bacterial Agents; D000903:Antibiotics, Antineoplastic; D000911:Antibodies, Monoclonal; D053281:Enediynes; C058024:C 1027