Development of and assay methodology for antibodies to benzo(a) pyrene (BP)

Journal Article · Thu May 01 00:00:00 EDT 1986 · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)

OSTI ID:5088875

Griffin, G D; Thomason, R; Murchison, C; Wecker, P; Kurka, K; Ambrose, K R

Rabbits, rats and mice have been immunized with BP-bovine serum albumin (BSA) conjugates, administered subcutaneously in Freund's adjuvant. Activity and specificity of antisera preparations from immunized animals was determined by: Double immunodiffusion, passive hemagglutination, enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay (RIA). All techniques could be successfully used to demonstrate the presence of BP antibodies in immune sera. The fist three assays were used only with BP antigen coupled to a protein. If BSA was used, cross-reactivity against BSA was observed, but differences in extent of reactivity toward BP-BSA and BSA were readily apparent. Antisera from highly immune animals showed anti-BP reactivity at 1-10 x 10/sup 6/ dilutions in the ELISA assay, but did not produce positive reactions in the passive hemagglutination assay at >1:256 dilutions. The RIA assay was used with either /sup 14/C-or /sup 3/H-BP, and relied upon an activated charcoal separation step to effectively remove (>98% efficiency) free BP from antibody-bound BP. Using this RIA assay, estimates of amounts of BP antibodies in antisera (0.1-1% of the immunoglobulin fraction) and antibody specificity for BP structural determinants could be made. These antibodies may form the basis for a novel detection system for BP and/or other polycyclic compounds.

• Research Organization: Oak Ridge National Lab., TN
• DOE Contract Number: AC05-84OR21400
• OSTI ID: 5088875
• Report Number(s): CONF-8606151-; TRN: 86-035183
• Journal Information: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Vol. 45:6; Conference: 76. annual meeting of the Federation of American Society for Experimental Biology, Washington, DC, USA, 8 Jun 1986
• Country of Publication: United States
• Language: English

Related Subjects

63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
59 BASIC BIOLOGICAL SCIENCES
BENZOPYRENE
RADIOIMMUNOASSAY
ANTIBODIES
CARBON 14 COMPOUNDS
MICE
RABBITS
RATS
TRITIUM COMPOUNDS
ANIMALS
AROMATICS
CONDENSED AROMATICS
HYDROCARBONS
IMMUNOASSAY
IMMUNOLOGY
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
MAMMALS
ORGANIC COMPOUNDS
RADIOASSAY
RADIOIMMUNOLOGY
RODENTS
TRACER TECHNIQUES
VERTEBRATES
560305* - Chemicals Metabolism & Toxicology- Vertebrates- (-1987)
550201 - Biochemistry- Tracer Techniques