Monoclonal antibodies to chlorinated dibenzo-p-dioxins

Stephen Kennel, G. Mason, S. Safe

Research output: Contribution to journalArticle

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. In order to screen the thousands of resultant cultures for production of MoAb, a rapid, solid phase radioimmunoassay for antibody to dioxins was developed. This 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 showed no significant binding to BSA-aniline while demonstrating high binding to BSA-TCDD (Ka = 4.5 × 1081 mole). Experiments are in progress to use these MoAbs to develop sensitive radioimmunoassays or enzyme linked assays specific for 2,3,7,8-TCDD.

Original languageEnglish (US)
Pages (from-to)2007-2010
Number of pages4
JournalChemosphere
Volume15
Issue number9-12
DOIs
StatePublished - Jan 1 1986

Fingerprint

Monoclonal antibodies
Aniline
dioxin
antibody
Bovine Serum Albumin
Monoclonal Antibodies
serum
Antibodies
Dioxins
Thyroglobulin
Hybridomas
Radioimmunoassay
Antigens
Assays
Polystyrenes
Cell Fusion
Fusion reactions
Enzymes
Enzyme Assays
fold

All Science Journal Classification (ASJC) codes

  • Environmental Science(all)
  • Environmental Chemistry

Cite this

Monoclonal antibodies to chlorinated dibenzo-p-dioxins. / Kennel, Stephen; Mason, G.; Safe, S.

In: Chemosphere, Vol. 15, No. 9-12, 01.01.1986, p. 2007-2010.

Research output: Contribution to journalArticle

Kennel, Stephen ; Mason, G. ; Safe, S. / Monoclonal antibodies to chlorinated dibenzo-p-dioxins. In: Chemosphere. 1986 ; Vol. 15, No. 9-12. pp. 2007-2010.
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