Immunological analysis of methamphetamine antibody and its use for the detection of methamphetamine by capillary electrophoresis with laser-induced fluorescence.
An accurate, simple and rapid immunoassay is demonstrated for the detection of methamphetamine in urine by capillary electrophoresis (CE) with laser-induced fluorescence (LIF). An aminobutyl derivative of methamphetamine was conjugated with proteins, and used as an immunogen to produce antibodies for the assay. The methamphetamine derivative was also labeled with fluorescein isothiocyanate (FITC) to compete with free methamphetamine in the sample for the antibody binding site. Levels of free and antibody-bound FITC-labeled methamphetamine were monitored by performing CE-LIF using an untreated fused-silica column. This competitive immunoassay used antiserum instead of purified antibody or antibody fragment, yet was found to have good precision with a sensitivity of lower than 20 ng/ml. Various antibodies were also screened, and cross-reactivity of anti-MA antibody with methamphetamine analogues were also investigated. The results indicate that CE-LIF-based immunoassay is a powerful tool for the screening and characterization of antibody and may have possible applications in the detection of abused drugs in urine.
Choi
J
J
Doping Control Center, Korea Institute of Science and Technology, Seoul, South Korea.
Kim
C
C
Choi
M J
MJ
D000818:Animals; D000937:Antigen-Antibody Reactions; D019075:Electrophoresis, Capillary / Q000379:methods; D016650:Fluorescein-5-isothiocyanate / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism*; D006041:Goats; D006801:Humans; D007106:Immune Sera / Q000378:metabolism* / Q000652:urine; D007118:Immunoassay; D007834:Lasers; D008694:Methamphetamine / Q000031:analogs & derivatives / Q000276:immunology* / Q000652:urine; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D005456:Fluorescent Dyes; D007106:Immune Sera; D008694:Methamphetamine; D016650:Fluorescein-5-isothiocyanate