ISSN:
1432-1335
Keywords:
Benzene
;
N 7-Phenylguanine
;
Monoclonal antibodies
;
Immunoassay
Source:
Springer Online Journal Archives 1860-2000
Topics:
Medicine
Notes:
Abstract N 7-Phenylguanine, a base adduct possibly formed after arylation of DNA by benzene oxide, the first reaction metabolite during benzene metabolism, was synthesized in our laboratory and used as reference for the production and characterization of monoclonal antibodies. 2-Hydroxymethyl-7-phenylhypoxanthine, a molecule structurally similar toN 7-phenylguanine, was coupled by a linker molecule to different carrier proteins. The resulting conjugate was used to immunize BALB/c mice, the spleen cells of which were fused with mouse P3X63-Ag8.653 myeloma cells to obtain monoclonal antibodies. Several hybridoma lines were cultivated in defined media and characterized as to sensitivity and specificity by an enzyme-linked immunosorbent assay (ELISA). Competitive ELISA demonstrated that all antibodies showed a very high affinity forN 7-phenylguanine but had a lower affinity towards various other samples includingN 7-chlorophenylguanines andC 8-,N 2- andO 6-phenylguanine. As little as about 20 pgN 7-phenylguanine could be detected with one of the most sensitive antibodies, CE6/G11, with a colorimetric end point while the detection limit could be lowered to about 10 pgN 7-phenylguanine when a fluorescent end point was used. The detection limit of other methods used to determineN 7-phenylguanine so far is 10 ng for gaschromatography/mass-spectrometry and 1 ng for high-pressure liquid chromatography. Thus the use of specific monoclonal antibodies seems to be the most sensitive method for the detection ofN 7-phenylguanine.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF01624434
Permalink
