title: Sensitive bioassay for detection of PPARα potentially hazardous ligands with gold nanoparticle probe.
abstract: There are so many kinds of peroxisome proliferator-activated receptor α (PPARα) ligands with hazardous effect for human health in the environment, such as certain herbicides, plasticizers and drugs. Among these agonists, perfluorooctane sulfonate (PFOS), perfluorooctanoic acid (PFOA), and mono-(2-ethylhexyl) phthalate (MEHP) are mostly investigated due to their persistence and accumulation in environment and their potential toxicity via PPARα. This investigation aims at developing a bioassay method to detect PPARα ligands based on the ligand-receptor interaction on microplate. PPARα, which formed heterodimers with retinoid X receptor-α (RXRα), were activated by PPARα ligands to form ligands-PPARα-RXRα complexes. Then the complexes were transferred into a microplate and captured via monoclonal anti-PPARα antibody. The PPARα responsive elements (PPRE) modified-gold nanoparticle probes were captured by the ligand-PPARα-RXRα complexes immobilized on the microplate, and then could be quantified through measuring the optical density after silver enhancement. The results showed that PFOS was quantified with a linear range from 100 pM to 1 μM and the detection limit was 10 pM. In addition to PFOS, PFOA and MEHP were also quantified within a proper range through the proposed bioassay. This bioassay was compared with that of liquid chromatography tandem-mass spectrometry (LC-MS) for water spiked samples with a significant correlation (r = 0.9893). This study provides a high-throughput detection method for PPARα ligands in microplate with high sensitivity and wide linear range. It may serve as an assistant of LC-MS for prescreening of PPARα ligands like PFOS.
authors: Minister of Education Key Laboratory of Environment and Health, School of Public Health, Tongji Medical College of Huazhong University of Science and Technology, Wuhan, Hubei Province 430030, PR China.
authors: Wei
authors: W
authors: Xia
authors: Yan-Jian
authors: YJ
authors: Wan
authors: Xianliang
authors: X
authors: Wang
authors: Yuan-Yuan
authors: YY
authors: Li
authors: Wen-Jie
authors: WJ
authors: Yang
authors: Chun-Xiang
authors: CX
authors: Wang
authors: Shun-qing
authors: SQ
authors: Xu
mesh_terms: D017738:Alkanesulfonic Acids / Q000737:chemistry; D000818:Animals; D001681:Biological Assay / Q000379:methods*; D002210:Caprylates / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004051:Diethylhexyl Phthalate / Q000031:analogs & derivatives / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D004784:Environmental Monitoring / Q000379:methods; D005466:Fluorocarbons / Q000737:chemistry; D006046:Gold / Q000737:chemistry*; D006801:Humans; D008024:Ligands; D008099:Liver / Q000378:metabolism*; D008297:Male; D053768:Metal Nanoparticles / Q000737:chemistry*; D036103:Nanotechnology / Q000379:methods*; D009841:Oligonucleotides / Q000235:genetics; D047493:PPAR alpha / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D018508:Water Purification / Q000379:methods
chemical_list: D017738:Alkanesulfonic Acids; D002210:Caprylates; D005466:Fluorocarbons; D008024:Ligands; D009841:Oligonucleotides; D047493:PPAR alpha; D006046:Gold; C023036:perfluorooctanoic acid; C076994:perfluorooctane sulfonic acid; D004051:Diethylhexyl Phthalate; C016599:mono-(2-ethylhexyl)phthalate