Enzyme-linked immunosorbent assay of neocarzinostatin chromophore (NCS-chr) by use of a monoclonal antibody against NCS-chr analog.
A monoclonal antibody against NCS-chr was prepared and characterized. Because of the instability of NCS-chr, chemically synthesized stable analog compound, termed PS, was used as a hapten of immunogen. The obtained antibody, termed APS, reacted with NCS-chr, but neither with NCS, NCS-polystyrene-maleic acid conjugate (SMANCS), nor UV-irradiated NCS-chr. Epitope analysis using the compounds that have a structure similar to PS showed that APS recognized the total structure, particularly cyclopenten moiety, of PS. These results suggest that APS recognizes the enediyne structure of NCS-chr. Next, the inhibition enzyme-linked immunosorbent assay (ELISA) for determination of NCS-chr was established. The standard curve showed that the microgram order of NCS-chr were accurately measurable by the established ELISA. Furthermore, it was revealed that the established ELISA was more sensitive than the antibiotic activity determination, termed Bio-assay. The established ELISA will be useful as a quantitative method of NCS-chr.
Mizugaki
M
M
Department of Pharmaceutical Sciences, Tohoku University Hospital, Sendai, Japan.
Itoh
K
K
Nakamura
H
H
Ishida
N
N
D000903:Antibiotics, Antineoplastic / Q000737:chemistry* / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D053281:Enediynes; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D009353:Zinostatin / Q000031:analogs & derivatives / Q000737:chemistry / Q000276:immunology
D000903:Antibiotics, Antineoplastic; D000911:Antibodies, Monoclonal; D053281:Enediynes; C032372:neocarzinostatin chromophore; D009353:Zinostatin