Production and characterization of antibody against aflatoxin Q1.
Antibodies against aflatoxin Q1 (AFQ1) were obtained from rabbits after immunization of either AFQ1-hemisuccinate or AFQ2a conjugated to bovine serum albumin. Both radioimmunoassay and enzyme-linked immunosorbent assaY (ELISA) were used for the determination of antibody titers and specificities. Antibodies obtained from rabbits after immunization with AFQ1-hemisuccinate-bovine serum albumin had the highest affinity to aflatoxin B1, whereas antibodies obtained from rabbits after immunization with AFQ2a-bovine serum albumin bound most effectively with AFQ2a. AFQ2a antibody was selected for the subsequent direct and indirect ELISA for the detection of AFQ1 in biological fluids. When AFQ2a-peroxidase and AFQ2a antibody were used, direct ELISA was able to detect as low as 2 ppb (ng/ml) of AFQ1 spiked in the urine samples that had been subjected to a Sep-Pak cleanup treatment. In indirect ELISA in which the antigen (AFQ2a-bovine serum albumin) was coated to the solid phase followed by reaction with rabbit antibody and goat anti-rabbit immunoglobulin G-peroxidase conjugate, 50-fold less antibody was used without sacrificing sensitivity. Recoveries of AFQ1 added to urine samples (2 to 40 ppb) were 46.3 to 73% and 65.8 to 85.8% for direct and indirect ELISA, respectively.
D000348:Aflatoxins / Q000276:immunology* / Q000652:urine; D000818:Animals; D000906:Antibodies / Q000032:analysis; D000917:Antibody Formation; D000918:Antibody Specificity; D004797:Enzyme-Linked Immunosorbent Assay; D011817:Rabbits; D011863:Radioimmunoassay; D012710:Serum Albumin, Bovine / Q000276:immunology
D000348:Aflatoxins; D000906:Antibodies; C041203:aflatoxin A1-hemisuccinate; C041205:aflatoxin Q2a-bovine serum albumin; D012710:Serum Albumin, Bovine; C033759:aflatoxin Q1; C041204:aflatoxin Q2a