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MONOCLONAL ANTIBODIES TO CHLORINATED DIBENZO-P-DIOXINS
Citation:
Kennel, S., G. Mason, AND S. Safe. MONOCLONAL ANTIBODIES TO CHLORINATED DIBENZO-P-DIOXINS. U.S. Environmental Protection Agency, Washington, D.C., EPA/600/J-86/536 (NTIS PB90165109), 1986.
Description:
A thyroglobulin conjugate of dioxin (thyroglobulin-2 adipamide, 3,7,8-trichlorodibenzo-p-dioxin) (TG-TCDD) was used to immunize BALB/c mice. ybridomas were produced by cell fusion between immune spleen cells and mouse myelomas SP2/0, P3, or 151. n order to screen the thousands of resultant cultures for production of MoAb, a rapid, solid phase radioimmunoassay for antibody to dioxins was developed. his 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. ourteen hybridomas were identified that produced MoAb reacting with ESA-TCDD but not with BSA alone. ntibodies were tested for binding to BSA-aniline to eliminate those with limited binding specificity. nitial studies indicated that while most MoAbs bound BSA-aniline as well as BSA-TCDD. ore 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. oAb 391-1B was purified from mouse ascites fluid and after radioiodination, was tested for direct binding to BSA-TCDD or BSA-aniline. 25I MoAb showed no significant binding to BSA-aniline wlile demonstrating high binding to ESA-TCDD (Ka = 4.5 x 10 8 1/mole). xperiments are in progress to use these MoAbs to develop senstive radioimmunoassays or enzyme linked assays specific for 2,3,7,8-TCDD.
