Mercapturic acids are urinary metabolites which signal exposure to genotoxic compounds. Carefully designed hapten-protein conjugates and a judicious screening strategy has enabled the generation of compound and class-specific antibodies which bind mercapturic acids. S-phenylmercapuric acid (S-PMA) is a highly specific and sensitive marker of benzene exposure. A monoclonal antibody reactive with S-PMA has been generated. The immobilised antibody retains immunoreactivity and can be used to enrich S-PMA from the urine of benzene exposed workers. The performance of the immunoaffinity column has been validated by comparison with data obtained from GC/MS analysis of urine from benzene exposed workers (range 12-168ug/1. corr. coeff. 0.98, n=23). Furthermore immunoaffinity chromatography facilitates the quantitative determination of urinary S-PMA by HPLC. Bioconcentration of S-PMA from the urine of benzene exposed workers has permitted the quantification of S-PMA by HPLC at 8 hour Time Weighted Average exposures of around 1ppm. Monoclonal antibodies to low molecular weight mercapturic acids (eg. S-(2-hydroxyethyl)mercapturic acid) were generally of too low affinity for practical application. As an alternative approach antibodies to adducted protein were investigated. Antibody 4D3 binds the adducted N-terminal heptapeptide released from the alpha-chain of haemoglobin by trypsin hydrolysis. Initial studies suggest antibody 4D3 can bind the adducted heptapeptide in whole haemoglobin. This may facilitate the determination of adducted haemoglobin in biomonitoring studies.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:641250 |
| Date | January 1997 |
| Creators | Ball, Lathan |
| Publisher | University of Edinburgh |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Source | http://hdl.handle.net/1842/10715 |
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