Fluorimetric immunoassay for multianalysis of aflatoxins

A sensitive fluorimetric elisa was developed for the analysis of aflatoxins. the assay was performed in a 384 microwell plate,wherein high specificity monoclonal antibody against afm1 (mab-afm1) was used as capture antibody and fitc conjugated secondary antibody was used for detection and quantification of the analyte. the linear range of the immunoassay was found to be 6.25-50 pg/ml. afm1 as low as 1 pg/ml was detected by this method with assay volume 40 l. the multi-analysis of different aflatoxins was also investigated in the microwell plate,based on the cross-reactivity (cr) approach. real milk samples were tested along with certified reference material by standard addition method and recovery analysis was done. the mab-afm1 showed 23.2% cr with afb1,50% cr with respect to afm2,and least cr towards afg1 (<1%). furthermore,mixture analysis of afm2 and afb1 was carried out at specific concentrations of afm1. the advantages of this developed immunoassay are high sensitivity,high throughput,multianalyte detection,versatility,and ease of handling. © 2013 lizy kanungo and sunil bhand.