Selection and Application of ssDNA Aptamers against Clenbuterol Hydrochloride Based on ssDNA Library Immobilized SELEX.
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, Kd, 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.
State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Synergetic Innovation Center of Food Safety and Nutrition, Jiangnan University , Wuxi 214122, China.
Nuo
N
Duan
Market Supervision and Administration Bureau , Taicang 215400, China.
Wenhui
W
Gong
State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Synergetic Innovation Center of Food Safety and Nutrition, Jiangnan University , Wuxi 214122, China.|School of Food and Biological Engineering, Jiangsu University , Zhenjiang 212013, China.
Shijia
S
Wu
State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Synergetic Innovation Center of Food Safety and Nutrition, Jiangnan University , Wuxi 214122, China.|National Engineering Research Center of Seafood, School of Food Science and Technology, Dalian Polytechnic University , Dalian 116034, China.
Zhouping
http://orcid.org/0000-0002-3868-8125
Z
Wang
D000318:Adrenergic beta-Agonists / Q000032:analysis*; D000818:Animals; D052157:Aptamers, Nucleotide / Q000737:chemistry* / Q000235:genetics; D001681:Biological Assay / Q000295:instrumentation / Q000379:methods*; D002976:Clenbuterol / Q000032:analysis*; D004277:DNA, Single-Stranded / Q000737:chemistry* / Q000235:genetics; D005506:Food Contamination / Q000032:analysis*; D008460:Meat / Q000032:analysis; D052156:SELEX Aptamer Technique / Q000295:instrumentation / Q000379:methods*; D013552:Swine
D000318:Adrenergic beta-Agonists; D052157:Aptamers, Nucleotide; D004277:DNA, Single-Stranded; D002976:Clenbuterol
SELEX; aptamer; clenbuterol hydrochloride; library immobilization