An immunological method for the detection of captopril-protein conjugate.
To clarify the mechanism of cytotoxicity induced by captopril (Cp), interactions between tissue protein and Cp were studied by the enzyme-linked immunosorbent assay (ELISA) method. The amide-linked adducts [hemocyanin from keyhole limpet-Cp adduct (KLH-Cp) and poly-L-Lys-Cp adduct (pLys-Cp)] using carbodiimide, and a disulfide-linked adduct [ovalbumin-Cp adduct (OVA-Cp)] were prepared. To determine the formation of the protein-Cp adduct, rabbit antisera against KLH-Cp was employed. The immunoreactivities with pLys-Cp and OVA-Cp against anti KLH-Cp sera were elevated when compared with the unmodified protein. With the inhibition ELISA, this antisera was useful for detecting the Cp disulfide-linked conjugate. In kidney cytosol, a high level of immunoreactivity was observed. Plasma and liver cytosol reactivities were similar, and approximately 40% against kidney cytosol. Thus, a method for the detection of the Cp-protein adduct using ELISA has been established. Formation of the Cp-protein adduct was observed in rat plasma and in liver and kidney cytosol. These findings suggest the possibility that the formation of Cp-protein adduct is partially related to cytotoxicity in liver.
Narazaki
R
R
Faculty of Pharmaceutical Sciences, Kumamoto University, Japan.
Watanabe
H
H
Maruyama
T
T
Suenaga
A
A
Otagiri
M
M
D000818:Animals; D002216:Captopril / Q000737:chemistry / Q000378:metabolism*; D003600:Cytosol / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006433:Hemocyanins / Q000737:chemistry; D007668:Kidney / Q000378:metabolism; D008099:Liver / Q000378:metabolism; D010047:Ovalbumin / Q000737:chemistry; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D051381:Rats
D011506:Proteins; D010047:Ovalbumin; D006433:Hemocyanins; D002216:Captopril