Record Display for the EPA National Library Catalog

RECORD NUMBER: 21 OF 30

Main Title Monoclonal Antibodies to Chlorinated Dibenzo-p-Dioxins.
Author Kennel, S. J. ; Mason, G. ; Safe, S. ;
CORP Author Oak Ridge National Lab., TN. Biology Div. ;Texas A and M Univ., College Station.;Environmental Research Lab.-Duluth, MN.
Publisher c1986
Year Published 1986
Report Number EPA-810995; EPA/600/J-86/536;
Stock Number PB90-265109
Additional Subjects Antigens ; Mice ; Reprints ; Monoclonal antibodies ; Trichlorodibenzodioxin ; Substrate specificity ; Hybridomas ; Radioimmunoassay ; Bovine serum albumin ; Binding sites
Holdings
Library Call Number Additional Info Location Last
Modified
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Status
NTIS  PB90-265109 Some EPA libraries have a fiche copy filed under the call number shown. 07/26/2022
Collation 5p
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/O, 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. (125)I MoAb showed no significant binding to BSA-aniline while demonstrating high binding to BSA-TCDD (Ka = 4.5 x 10 to the eighth/mole).