14 The Development and Characterisation of Novel Monoclonal Antibodies (Mabs) for 1-Hydroxypyrene Glucuronide (1-OHPyrG) Determination and Polycyclic Aromatic Hydrocarbon (PAH) Biomonitoring

Abstract

Abstract Objective PAHs are produced during the incomplete burning of organic materials. PAH sources include vehicle exhaust, tobacco smoke and waste incineration. Cancer is a significant endpoint of PAH exposure. Pyrene is a common component of PAH exposures and metabolism of pyrene leads to the excretion of 1-OHPyrG in urine. Measurement of 1-OHPyrG has been used to assess occupational and environmental exposure to PAHs. The production of Mabs to 1-OHPyrG will allow the development of novel PAH biomonitoring tests, facilitating the introduction of routine screening and safeguarding human health. Methods 1-Pyrenebutyric acid was coupled to a carrier protein and a sheep Mab raised to 1-OHPyrG. A second Mab which binds the Mab-1-OHPyrG immune complex was also produced and a novel urinary non-competitive ELISA developed. Careful formulation of assay buffers has overcome the heterogeneous nature of urine. Assay performance was confirmed through binding studies with structurally similar metabolites, reproducibility testing and a comparative method study. Results A sensitive and specific ELISA (measuring range 20-500pg/ml) was used to detect elevated levels of 1-OHPyrG in urine. Occupational samples determined by ELISA and HPLC-fluorescence were in good agreement (R2=0.91, N=20). A test kit is now being transferred to manufacture. Conclusions The development of a novel ELISA for 1-OHPyrG has allowed the production of a sensitive and specific PAH biomonitoring test kit. Kits increase the accessibility of biomonitoring and facilitate the introduction of routine screening. Alternate assay formats are being investigated. A lateral flow “point of care” test will enable on-site testing and significantly increase the utility of biomonitoring.