title: General Bioluminescence Resonance Energy Transfer Homogeneous Immunoassay for Small Molecules Based on Quantum Dots.
abstract: Here, we describe a general bioluminescence resonance energy transfer (BRET) homogeneous immunoassay based on quantum dots (QDs) as the acceptor and Renilla luciferase (Rluc) as the donor (QD-BRET) for the determination of small molecules. The ratio of the donor-acceptor that could produce energy transfer varied in the presence of different concentrations of free enrofloxacin (ENR), an important small molecule in food safety. The calculated Förster distance (R0) was 7.86 nm. Under optimized conditions, the half-maximal inhibitory concentration (IC50) for ENR was less than 1 ng/mL and the linear range covered 4 orders of magnitude (0.023 to 25.60 ng/mL). The cross-reactivities (CRs) of seven representative fluoroquinolones (FQs) were similar to the data obtained by an enzyme-linked immunosorbent assay (ELISA). The average intra- and interassay recoveries from spiked milk of were 79.8-118.0%, and the relative standard deviations (RSDs) were less than 10%, meeting the requirement of residue detection, which was a satisfactory result. Furthermore, we compared the influence of different luciferase substrates on the performance of the assay. Considering sensitivity and stability, coelenterazine-h was the most appropriate substrate. The results from this study will enable better-informed decisions on the choice of Rluc substrate for QD-BRET systems. For the future, the QD-BRET immunosensor could easily be extended to other small molecules and thus represents a versatile strategy in food safety, the environment, clinical diagnosis, and other fields.
authors.affiliation: Beijing Advanced Innovation Center for Food Nutrition and Human Health, College of Veterinary Medicine, China Agricultural University , No. 2 Yuanmingyuan West Road, Beijing 100193, China.
authors.forename: Xuezhi
authors.initials: X
authors.lastname: Yu
authors.affiliation: Beijing Advanced Innovation Center for Food Nutrition and Human Health, College of Veterinary Medicine, China Agricultural University , No. 2 Yuanmingyuan West Road, Beijing 100193, China.
authors.forename: Kai
authors.initials: K
authors.lastname: Wen
authors.affiliation: Beijing Advanced Innovation Center for Food Nutrition and Human Health, College of Veterinary Medicine, China Agricultural University , No. 2 Yuanmingyuan West Road, Beijing 100193, China.
authors.forename: Zhanhui
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Beijing Advanced Innovation Center for Food Nutrition and Human Health, College of Veterinary Medicine, China Agricultural University , No. 2 Yuanmingyuan West Road, Beijing 100193, China.
authors.forename: Xiya
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Beijing Advanced Innovation Center for Food Nutrition and Human Health, College of Veterinary Medicine, China Agricultural University , No. 2 Yuanmingyuan West Road, Beijing 100193, China.
authors.forename: Chenglong
authors.initials: C
authors.lastname: Li
authors.affiliation: Beijing Advanced Innovation Center for Food Nutrition and Human Health, College of Veterinary Medicine, China Agricultural University , No. 2 Yuanmingyuan West Road, Beijing 100193, China.|Beijing Laboratory for Food Quality and Safety and Beijing Key Laboratory of Detection Technology for Animal-Derived Food Safety , No. 2 Yuanmingyuan West Road, Beijing 100193, China.
authors.forename: Suxia
authors.initials: S
authors.lastname: Zhang
authors.affiliation: Beijing Advanced Innovation Center for Food Nutrition and Human Health, College of Veterinary Medicine, China Agricultural University , No. 2 Yuanmingyuan West Road, Beijing 100193, China.|Beijing Laboratory for Food Quality and Safety and Beijing Key Laboratory of Detection Technology for Animal-Derived Food Safety , No. 2 Yuanmingyuan West Road, Beijing 100193, China.
authors.forename: Jianzhong
authors.initials: J
authors.lastname: Shen
mesh_terms: D060807:Bioluminescence Resonance Energy Transfer Techniques; D000077422:Enrofloxacin; D024841:Fluoroquinolones / Q000032:analysis* / Q000378:metabolism; D007118:Immunoassay; D049410:Luciferases, Renilla / Q000737:chemistry / Q000378:metabolism; D045663:Quantum Dots
chemical_list: D024841:Fluoroquinolones; D000077422:Enrofloxacin; D049410:Luciferases, Renilla