Journal
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
Volume 65, Issue 8, Pages 1771-1777Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.jafc.6b04951
Keywords
aptamer; clenbuterol hydrochloride; library immobilization; SELEX
Funding
- Key Research and Development Program of Jiangsu Province [BE2016306]
- National Science and Technology Support Program of China [2015BAD17B02]
- Natural Science Foundation of Jiangsu Province [BK201401SS]
- National Natural Science Foundation of China (NSFC) [31401575]
- China Postdoctoral Science Foundation [2016T90430]
- Collaborative innovation center of food safety and quality control in Jiangsu Province
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Clenbuterol hydrochloride (CLB) is often abused as additive feed for livestock to decrease adipose tissue deposition and to increase growth rate. It raises a potential risk to human health through the consumption of animal product. In this study, aptamers with higher affinity and specificity were screened through 16 selection rounds based on the ssDNA library immobilized systematic evolution of ligands by exponential enrichment (SELEX) technique. After cloning and sequencing, five aptamer candidates were picked out for affinity and specificity assays based on a graphene oxide (GO) adsorption method. The results showed that the aptamer CLB-2 binds specifically against CLB with a dissociation constant, IC, value of 76.61 12.70 nM. In addition, an aptamer-based fluorescence bioassay was established for CLB analysis. The correlation between the CLB concentration and fluorescent signal was found to be linear within the range of 0.10 to 50 ng/mL with a limit of detection of 0.07 ng/mL. It has been further applied for the determination of CLB in pork samples, showing its great potential for sensitive analysis in food safety control.
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