PREPARATION AND CHARACTERIZATION OF AFLATOXIN B_2a‐HEMIGLUTARATE AND ITS USE FOR THE PRODUCTION OF ANTIBODY AGAINST AFLATOXIN B₁

Abstract

Hemisuccinate (HS) and hemiglutarate (HG) of aflatoxin B2a (afla B2a) were prepared by refluxing afla B2a with the corresponding anhydride and 4‐N, N‐dimethylaminopyridine in tetrahydrofuran. Two epimers of the respective HS or HG which show different chromatographic behavior and physiochemical properties were isolated and characterized. Afla B2a‐HS hydrolyzes very rapidly in aqueous solution and was not used for further study. Afla B2a‐HG hydrolyzes at a much slower rate and was selected for the coupling to protein. Using the mixed anhydride method, as much as 12 moles of afla B2a‐HG were conjugated to each mole of bovine serum albumin (BSA). The antibody obtained from rabbits immunized with afla B2a‐HG BSA is most specific to afla B1 and shows little cross reaction with afla G1 and aflatoxicol. The lower limit for detection of afla B1 by radioimmunoassay using this antibody is in the range of 30–50 pg per assay.

Production and characterization of antibody against diacetoxyscirpenol

“…The effectiveness of HG derivatives of mycotoxins as haptens has been demonstrated in two previous studies (8). The HG of aflatoxin B2a was also demonstrated to be a good immunogen for the production of antibody against aflatoxin B2 (8).…”

A morphine/heroin vaccine with new hapten design attenuates behavioral effects in rats

“…We had three reasons for choosing this structure for the hapten. First, aflatoxin B2a‐hemisuccinate hydrolyzes very rapidly in aqueous solution, whereas aflatoxin B2a‐hemiglutarate hydrolyzes at a much slower rate (Lau et al. 1980), which may stabilize the vaccine with the 6‐glutaratemorphine hapten to trigger a sustained and robust humoral immunologic response.…”

Indirect enzyme-linked immunosorbent assay for the mycotoxin zearalenone

“…Since zearalenone has no reactive group for coupling reactions, it was first converted to zearalenone-6'-carboxymethyloxime (Z-oxime) by the method of Thouvenot and Morfin (26). Z-oxime was then conjugated to BSA by a mixed anhydride procedure for use as an immunogen (15). Briefly, Z-oxime (15 mg) was dissolved in 5 ml of dry tetrahydrofuran and cooled to -5°C in an ice-salt bath.…”