Abstract:
ObjectiveTo prepare monoclonal antibody against aflatox in M
1 and develop indirect competitive enzymelinked immunosorbent assay for detection of aflatox in M
1.
MethodHybridoma cell line excreting monoclonal antibody against aflatox in M
1 was produced using B cell hybridoma technique and develop indirect competitive enzyme-linked immunosorbent assay for detection of aflatox in M
1.
ResultsOne hy bridoma cell line excreting monoclonal antibodies against aflatox in M
1 coded 2F2 was obtained by fusing murine Sp2/0 cells with spleen cells from BALB/c mice immunized with A FM
1-BSA conjugate.The monoclonal antibody produced by the hybridoma cell were tested for subtype and designated as IgG
1 for 2F2.The affinity of IgG in purified ascites yielded by hybridoma cell was 2.8×10
-11 mol/L.The monoclonal antibody obtained in the present study was specific to aflatoxin M
1,because of lightly cross reactions among the monoclonal antibodies against aflatox in M
1 with the analog ues of aflatox in B
1,aflatoxin B
2,aflatox in G
1,aflatox in G
2 and aflatox in M
2 were found.The limit of detect ing concentr at ion of aflatox in M
1 was 0.07 ng/ml.The linear range of standard curve was 0.02~2 ng/ml and the linearequation was y=-0.436 4x+0.269 3(
R2=0.994 9).The recovery of aflatox in M
1 was betw een 72.5% ~131.3%.
ConclusionThe monoclonal antibody obtained in the study was of relatively high specificity to aflatox in M
1,and a simple fast sensitive indirect competitive enzyme-linked immunosorbent assay for the detection of aflatox in M
1 was developed.