Synthesis of Novel Haptens and Conjugates for Antibody Production against Kainoid Family

Abstract

Amnesic shellfish poisoning (ASP) is caused by consumption of contaminated seafood that has accumulated kainic acid or kainoid analogues such as domoic acid. Among the different ASP bioassays, immunoassays are an attractive alternative to the in vivo mouse bioassay. Herein, we report the synthesis and bioconjugation of two new haptens for the generation of a specific antibody against members of the kainoid family.

ASP - kainoids - haptens - antibody - ELISA

References and Notes

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(2 S ,3 S )-Dibenzyl 3-[2-(Benzyloxy)-2-oxoethyl]-4-[4-( tert -butoxy)-4-oxobut-2-en-1-ylidene]pyrrolidine-1,2-dicarboxylate (12): The same experimental procedure as described for hapten 8 was used for compound 12 (yield: 80%). ^¹H NMR (200 MHz, CDCl₃): δ (mixture of stereo-isomers and conformers) = 2.56-2.84 (m, 2 H), 3.31-3.52 (m, 0.7 H), 3.65-3.90 (m, 0.3 H), 4.32-4.57 (m, 3 H), 5.01-5.23 (m, 6 H), 5.79 (d, J = 14.9 Hz, 1 H), 6.07 (d, J = 12.2 Hz, 1 H), 7.18-7.42 (m, 16 H), 10.00 (br s, 1 H). ^¹³C NMR (50 MHz, CDCl₃): δ (mixture of stereoisomers and conformers) = 38.9 (CH₂), 44.9, 45.8 (CH), 48.3, 48.9 (CH₂), 63.7 (CH), 66.5-67.8 (CH₂), 121.9 (CH), 128.4-128.7 (CH), 135.4, 136.0 (C), 141.3 (CH), 147.7, 148.7 (C), 154.7, 155.2 (C), 170.2-171.4 (C). HRMS (ES): m/z [M + Na⁺] calcd for C₃₃H₃₁NO₈Na: 592.1941; found: 592.1926.