Monoclonal antibodies to chlorinated dibenzo-p-dioxins.
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abstract
A thyroglobulin conjugate of dioxin (thyroglobulin-2 adipamide, 3,7,8-trichlorodibenzo-p-dioxin) (TG-TCDD) was used to immunize BALB/c mice. Hybridomas were produced by cell fusion between immune spleen cells and mouse myelomas SP2/0, P3, or NS1. To screen the thousands of resultant cultures for production of monoclonal antibodies (MoAb), a rapid, solid-phase radioimmunoassay for antibody to dioxins was developed. This procedure involved attaching bovine serum albumin coupled with trichlorodibenzo-p-dioxin (BSA-TCDD) to polystyrene plates to be used as a solid-phase target antigen for reaction with MoAb. Fourteen hybridomas were identified that produced MoAb reacting with BSa-TCDD but not with BSA alone. Antibodies were tested for binding to BSA-aniline to eliminate those with limited binding specificity. Initial studies indicated that most MoAbs bound BSA-aniline as well as BSA-TCDD. More detailed analyses indicated that while most MoAbs showed some reaction with BSA-aniline, two showed preferential binding to BSA-TCDD of more than 200-fold whereas rabbit antisera demonstrated only a 5-fold discrimination. MoAb 391-1B was purified from mouse ascites fluid and after radioiodination, was tested for direct binding to BSA-TCDD or BSA-aniline. 125I-MoAb 391-1B showed no significant binding to BSA-aniline while demonstrating high binding to BSA-TCDD (Ka = 4.5 X 10(8) liters/mol).
