Immunoassay for methamphetamine with a new antibody.
p- and o-Aminomethamphetamine were synthesized as haptens to be coupled with carrier protein at the benzene ring of methamphetamine. Immunogens were prepared by the glutaraldehyde method or the MBS (N-(m-maleimidobenzoyloxy)succinimide) type cross-linking reagent method. In particular, immunization with p-aminomethamphetamine-bovine serum albumin (BSA) conjugate prepared by the glutaraldehyde method gave an anti-methamphetamine antiserum having a low cross-reactivity with methylephedrine. With the antiserum, three kinds of immunoassays for methamphetamine were established. An enzyme immunoassay (EIA) and an enzyme-linked immunosorbent assay (ELISA) were developed with alkaline phosphatase (ALP) as a label enzyme. The amount of antibody bound ALP conjugate was determined by its activity in dephosphorylating p-nitrophenyl phosphate in EIA and nicotinamide adenine dinucleotide phosphate (NADP+) in ELISA. The range of methamphetamine measurable by ELISA was 0.025-0.5 ng/well and its sensitivity was superior to that of EIA (0.3-300 ng/tube). A latex agglutination inhibition reaction test (LAIRT) was also developed for the mass screening method of urine samples. The sensitivity of this method for methamphetamine was 0.1 micrograms/ml urine.
Aoki
K
K
Department of Biochemical Toxicology, Pharmaceutical Sciences, Showa University, Tokyo, Japan.
Hirose
Y
Y
Kuroiwa
Y
Y
D000469:Alkaline Phosphatase; D000818:Animals; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D006241:Haptens / Q000138:chemical synthesis; D006801:Humans; D007106:Immune Sera / Q000276:immunology; D007124:Immunoenzyme Techniques; D007841:Latex Fixation Tests; D008694:Methamphetamine / Q000032:analysis* / Q000276:immunology / Q000652:urine; D011237:Predictive Value of Tests; D011817:Rabbits; D014614:Vaccines, Synthetic
D006241:Haptens; D007106:Immune Sera; D014614:Vaccines, Synthetic; D008694:Methamphetamine; D000469:Alkaline Phosphatase