Determination of phenanthrene by antibody-coated competitive real-time immuno-PCR assay |
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Authors: | Chun Zhou Qiong-E Wang Hui-Sheng Zhuang |
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Institution: | (1) College of Environmental Science and Engineering, DongHua University, 201620 Shanghai, China;(2) College of Chemical Engineering, DongHua University, 201620 Shanghai, China |
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Abstract: | A reliable selective and sensitive antibody-coated competitive real-time immuno-PCR (RT-IPCR) assay for the determination
of phenanthrene (PH) was developed. Phenanthrene butanoic acid (γ-oxo-PHA) was synthesized as the hapten of PH. An active
ester method was used to couple the PHA to bovine serum albumin to form an artificial immune antigen. Male New Zealand white
rabbits were immunized with immune antigen to obtain polyclonal antibodies, with which a novel RT-IPCR assay for determination
of PH was developed. Under the optimized assay conditions, PH can be determined in the concentration range from 10 fg/mL to
100 pg/mL with a detection limit of 5 fg/mL. The cross-reactivities of the anti-PH antibody to seven structurally related
compounds were below 12.5%. Some environmental water samples were analyzed with satisfactory results, which showed good accuracy
and suitability to analyze PH in environmental water. Compared with high-performance liquid chromatography, the recovery was
lower or higher with agitation but would still be acceptable for use in an on-site field test to provide rapid, semiquantitative,
and reliable test results for making environmental decisions. |
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Keywords: | Phenanthrene Antigen Polyclonal antibody Real-time immuno-PCR assay |
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