Study On Determination Of Fumonisin B₁ By Immunoassay And Immunoaffinity Column

In this paper, two anti-FB₁-polyclonal antibodies were produced by a complete FB₁ antigen synthesised by FB₁ and KLH through glutaraldehyde. Based on the affinity of these two antibody,No. 1 antibody was chosen for future use.Under optimized conditions of the FB₁-ELISA test, the test limit achieved as low as 0.18±0.08μg/kg. Corn, barley, wheat and oats were used as real samples (negative sample detected by HPLC) for analysis FB₁ and their matrix effect on ELISA can be eliminated by diluting and adding enshrouding reagents. Then the recovery test showed that all the test of the above four kinds of crop could reach a recovery range between 65% to 95%. The stability of both antibody and enzyme liked antigen of FB₁-ELISA been tested for the development of FB₁-ELISA test kit. The results proved that both of antibody and enzyme linked FB₁ were stable under 4℃for 6 months.The study of FB₁-ELISA test with immunoaffinity column purification started based on plenteous antibody storage. The immuoaffmity columns were prepared by coupling anti-FB₁multiclonal antibody and OVA with sepharose-4B. High coupling rates were determined by UV absorbance measurement. Then the coulumn operating conditions (elution buffer, elution volume, washing buffer, enshrouding reagents et. al) were optimized through direct-competetion ELISA. Then the purification capacity of the FB₁-IAC was tested by using corn as a sample. Result of the test showed that immunoaffinity column could successfully eliminate the impact corn brings to ELISA. The mean recoveries were in the range 52%-63%.