Development of a liquid phase radioimmunoassay technique for the detection of aflatoxin B1

Abstract EN

This study aimed to prepare the basic components of aflatoxin B1 (AfB1) radioimmunoassay (RIA) technique using liquid phase to measureAfB1 in agricultural commodities.

These components were polyclonal antibody (anti-AfB1), 125I-labelledAfB1 and aflatoxin B1 standards.Anti-AfB1 polyclonal was raised by immunizing six white New-Zealand rabbits with aflatoxin B1- bovine serum albumin (AfB1-BSA) conjugate as immunogen after four months through primary injection and three boosters via subcutaneous and intramuscular routes.AfB1 oxime derivative was synthesized, characterized, and used for preparation of 125I-labeled AfB1 tracer.

Preparation of AfB1 standards was carried out using highly purified AfB1antigen in ethanol as a matrix.

The obtained AfB1-antisera were characterized in terms of immunoresponse and titer which gave an optimum dilution of 1:1000.

Optimization and validation of the local liquid phase RIA system were carried out.

The results obtained provide a valid, accurate and highly sensitive RIA system of AfB1 reaching to 0.28 ng/ ml.

In conclusion, this assay could be used in laboratories to measure low levels of AfB1 in agricultural commodities to avoid prolonged exposure to sub-acute levels, which can lead to hepatotoxicity and liver cancer in animals and humans.