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Toxins (Basel). 2016 May 12;8(5). pii: E148. doi: 10.3390/toxins8050148.

Monoclonal IgA Antibodies for Aflatoxin Immunoassays.

Author information

1
Genetic Engineering and Biotechnology Institute, Marmara Research Center, The Scientific and Technological Research Council of Turkey, TÜBİTAK, Gebze, Kocaeli 41400, Turkey. ozlem.ertekin@tubitak.gov.tr.
2
Department of Molecular Biology and Genetics, Graduate School of Sciences, Gebze Technical University, Gebze, Kocaeli 41400, Turkey. ozlem.ertekin@tubitak.gov.tr.
3
Genetic Engineering and Biotechnology Institute, Marmara Research Center, The Scientific and Technological Research Council of Turkey, TÜBİTAK, Gebze, Kocaeli 41400, Turkey. seyda.pirincci@tubitak.gov.tr.
4
Department of Medical Genetics and Molecular Biology, Kocaeli University, Umuttepe, Kocaeli 41100, Turkey. seyda.pirincci@tubitak.gov.tr.
5
Genetic Engineering and Biotechnology Institute, Marmara Research Center, The Scientific and Technological Research Council of Turkey, TÜBİTAK, Gebze, Kocaeli 41400, Turkey. selma.ozturk@tubitak.gov.tr.

Abstract

Antibody based techniques are widely used for the detection of aflatoxins which are potent toxins with a high rate of occurrence in many crops. We developed a murine monoclonal antibody of immunoglobulin A (IgA) isotype with a strong binding affinity to aflatoxin B1 (AFB1), aflatoxin B2 (AFB2), aflatoxin G1 (AFG1), aflatoxin G2 (AFG2) and aflatoxin M1 (AFM1). The antibody was effectively used in immunoaffinity column (IAC) and ELISA kit development. The performance of the IACs was compatible with AOAC performance standards for affinity columns (Test Method: AOAC 991.31). The total binding capacity of the IACs containing our antibody was 111 ng, 70 ng, 114 ng and 73 ng for AFB1, AFB2, and AFG1 andAFG2, respectively. Furthermore, the recovery rates of 5 ng of each AF derivative loaded to the IACs were determined as 104.9%, 82.4%, 85.5% and 70.7% for AFB1, AFB2, AFG1 and AFG2, respectively. As for the ELISA kit developed using non-oriented, purified IgA antibody, we observed a detection range of 2-50 µg/L with 40 min total test time. The monoclonal antibody developed in this research is hitherto the first presentation of quadruple antigen binding IgA monoclonal antibodies in mycotoxin analysis and also the first study of their utilization in ELISA and IACs. IgA antibodies are valuable alternatives for immunoassay development, in terms of both sensitivity and ease of preparation, since they do not require any orientation effort.

KEYWORDS:

ELISA; immunoaffinitycolumn; immunoglobulin A; monoclonal antibody; mycotoxin; orientation

PMID:
27187470
PMCID:
PMC4885063
DOI:
10.3390/toxins8050148
[Indexed for MEDLINE]
Free PMC Article

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