Choi, J Choi, MJ Kim, C Cho, YS Chin, J Jo, YA 2024-01-21T18:40:11Z 2024-01-21T18:40:11Z 2021-09-05 1997-02 0253-6269 https://pubs.kist.re.kr/handle/201004/143965 To obtain more sensitive immunoassay for methamphetamine (MA) determination, the optimum condition of enzyme-linked immunosorbent assay (ELISA) was investigated in regard to immunogens, antibody purification methods and coating tracers. Activated MA, N-(4-aminobutyl)methamphetamine (4-ABMA), was conjugated with bovine serum albumin (BSA) or keyhole limpet hemocyanin (KLH) and used as immunogen. The antibodies were purified by protein G chromatography or various immunoaffinity chromatography-linked MA-protein ligands, such as MA-BSA, MA-KLH or MA-ovalbumin (OVA). Each purified antibody was characterized by means of sensitivity and cross-reactivity using the three MA-protein coating tracers, MA-BSA, MA-KLH and MA-OVA. The best sensitivity of each antibody was acquired with the MA-OVA tracer although the tracer concentration and the antibody titer level at optimum condition were varied. The antibody with high titer level did not always yield good sensitivity. At optimum condition, immunoaffinity chromatography-purified antibodies were better for sensitivity and for specificity than protein C-purified antibodies. The cross-reactivity of the purified antibodies seemed to be affected by immunogen structure and showed somewhat different patterns according to the immunoaffinity ligand utilized. These data show that the antibody purification method as well as choice of coating tracer and immunogen is essential for the sensitivity and specificity of EIA; the optimum condition for assay should be discovered using various methods and combinations. English PHARMACEUTICAL SOCIETY KOREA POLARIZATION FLUOROIMMUNOASSAY AMPHETAMINE IMMUNOASSAY URINE The optimization of ELISA for methamphetamine determination: The effect of immunogen, tracer and antibody purification method on the sensitivity Article 10.1007/BF02974041 1 ARCHIVES OF PHARMACAL RESEARCH, v.20, no.1, pp.46 - 52 ARCHIVES OF PHARMACAL RESEARCH 20 1 46 52 scie scopus A1997WK36600009 2-s2.0-0002812204 Chemistry, Medicinal Pharmacology & Pharmacy Pharmacology & Pharmacy Article POLARIZATION FLUOROIMMUNOASSAY AMPHETAMINE IMMUNOASSAY URINE methamphetamine ELISA antibody purification method coating tracer optimization