Production of monoclonal antibody and development of enzyme-linked immunosorbent assay for kanamycin in biological matrices.
Monoclonal antibodies (MAbs) against kanamycin were prepared by using a kanamycin-bovine gamma-globulin conjugate for the immunization of mice. Splenocytes from BALB/c immunized mice were fused with P3X63Ag8U.1 myeloma cells. This resulted in two hybridoma cell lines. Fifty per cent inhibition concentrations (IC50) for the MAbs were 2 and 5 ng ml-1. One MAb (IC50 = 2 ng ml-1) was named #22 and was used to develop quantitative assays for kanamycin by means of an enzyme-linked immunosorbent assay (ELISA). The detection limit was 0.2 ng ml-1 and the standard deviations were 0.2-4.4% for intra-assay and 0.6-4.7% for inter-assay, respectively. The detection limits using peroxidase were 4 ppb in cattle milk, cattle plasma, cattle urine, swine plasma, swine urine and chicken plasma. Using the MAb #22 produced, a rapid test kit based on an immunochromatographic method was developed. The detection limits using the kit were 50 ppb in cattle milk, cattle plasma, cattle urine and chicken plasma.
Watanabe
H
H
Research Institute for Animal Science in Biochemistry and Toxicology, Kanagawa, Japan.
Satake
A
A
Kido
Y
Y
Tsuji
A
A
D000818:Animals; D000904:Antibiotics, Antitubercular / Q000032:analysis* / Q000097:blood / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D002645:Chickens; D004350:Drug Residues / Q000032:analysis*; D004797:Enzyme-Linked Immunosorbent Assay; D007612:Kanamycin / Q000032:analysis* / Q000097:blood / Q000276:immunology; D008892:Milk / Q000737:chemistry
D000904:Antibiotics, Antitubercular; D000911:Antibodies, Monoclonal; D007612:Kanamycin