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Immunological Detection of Aflatoxin B₁-DNA Adducts Formed in Vivo¹

Cancer Center and Division of Environmental Science, Columbia University, New York, New York 10032 [L-L. H., S-W. H., R. M. S.], and Graduate Institute of Clinical Medicine, National Taiwan University Hospital, Taiwan, Republic of China [D-S. C.]

Two monoclonal antibodies (6A10 and 12F5) were obtained after fusion of mouse P3X63-AG.8.653 myeloma cells with spleen cells isolated from BALB/c mice immunized with imidazole ring-opened aflatoxin B₁ (AFB₁)-DNA and characterized by competitive enzyme-linked immunosorbent assays. Both antibodies are highly specific for imidazole ring opened AFB₁-DNA and show some cross-reactivity with AFB₁-DNA and no cross-reactivity with 8,9-dihydro-8-(7-guanyl)-9-hydroxy-AFB₁, AFB₁ conjugated with bovine serum albumin, aflatoxin M₁ conjugated with bovine serum albumin, AFB₁, or aflatoxin G₁. Antibody 6A10 was further characterized and showed no cross-reactivity with DNA modified by several other carcinogens. It could detect adducts with 4-fold higher sensitivity in highly modified DNA (2.5 adducts/100 nucleotides) than in low modified DNA (4 adducts/10⁵ nucleotides). With low modified DNA the limit of sensitivity is 5 adducts/10⁷ nucleotides. Antibody 6A10 reliably detected adducts formed in vivo in rats and mice treated with AFB₁. In a pilot study, AFB₁ adducts were detected in liver tissues from individuals living in areas with suspected exposure to AFB₁. Monitoring adduct levels in human tissue may provide information not only on carcinogen exposure but also on the relationship among infection with hepatitis B virus, dietary exposure to aflatoxin B₁, and liver cancer.

¹ This work was supported by National Institute of Environmental Health Sciences Grant ESO 3881 and NIH Grant CA 21111.

² To whom requests for reprints should be addressed, at Division of Environmental Sciences, Columbia University, 650 W. 168th Street, New York, NY 10032.

Received 5/25/88. Revised 8/ 9/88. Accepted 8/15/88.

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