TY - JOUR
T1 - Monoclonal antibodies to chlorinated dibenzo-p-dioxins
AU - Kennel, Stephen J.
AU - Jason, Casey
AU - Albro, Phillip W.
AU - Mason, Grant
AU - Safe, Stephen H.
N1 - Funding Information:
’ Research sponsored by the Office of Health and Environmental Research, U.S. Department of Energy, under Contract DE-ACO5-840R2 1400 with the Martin Marietta Energy Systems,I nc. The U.S. Government’s right to retain a nonexclusive royalty-free license in and to the copyright covering this paper, for governmental purposes, is acknowledged. * To whom reprint requests should be addressed.
Copyright:
Copyright 2014 Elsevier B.V., All rights reserved.
PY - 1986/2
Y1 - 1986/2
N2 - 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 solidphase 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 × 108liters/mol).
AB - 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 solidphase 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 × 108liters/mol).
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U2 - 10.1016/0041-008X(86)90200-0
DO - 10.1016/0041-008X(86)90200-0
M3 - Article
C2 - 3945952
AN - SCOPUS:0022634504
SN - 0041-008X
VL - 82
SP - 256
EP - 263
JO - Toxicology and Applied Pharmacology
JF - Toxicology and Applied Pharmacology
IS - 2
ER -