Development of an Enzyme-Linked Immunosorbent Assay for 1-Nitropyrene

Abstract

An enzyme-linked immunosorbent assay (ELISA) was developed for 1-nitropyrene. For hapten syntheses pyrenebutyric acid was nitrated yielding a mixture of 4-(6-nitropyrene-1-y1)-and 4-(8-nitropyrene-1 -y1)-butyric acid isomers that could not be separated. High titered antisera were obtained in rabbits after immunization with a KLH immunogen prepared with the mixture of isomers. As coating antigen a corresponding BSA conjugate was used. The calibration curve was linear in the 0.1 to 10 ppb range with a center point at 1.5 ppb and an LOD of 0.02 ppb in 10:90 acetonitrile/water (v/v). The average coefficients of variation were 4.9 and 7.8% over the range of the standard curve for intra-and interassay precision. Interference caused by 1,6-and 1,8-dinitropyrenes was observed. The effect of different concentrations of solvents on the antibody-analyte interaction was studied.

Monoclonal Antibody-Based ELISAs for Part-per-Billion Determination of Polycyclic Aromatic Hydrocarbons: Effects of Haptens and Formats on Sensitivity and Specificity

“…These include extraction of the sample by either liquid−liquid or solid-phase extraction, concentration and cleanup of the extracts followed by analysis with high-performance liquid chromatography (HPLC), gas chromatography (GC) or GC/mass spectrometry (GC/MS) . To avoid these complex procedures, immunochemical analyses employing either polyclonal or monoclonal antibodies have been developed for PAHs. − Commercially available immunoassay kits for PAHs, total petroleum hydrocarbons (TPHs), and carcinogenic PAHs in various matrixes are marketed by Strategic Diagnostics, Inc. (Newark, DE). U.S. EPA has compiled a series of immunochemical methods for environmental analysis.…”