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Synthesis of Cd-free InP/ZnS Quantum Dots Suitable for Biomedical Applications
Published on: February 6, 2016
Application of CdTe/CdS/ZnS quantum dot in immunoassay for aflatoxin B1 and molecular modeling of antibody
Fuyuan Zhang1, Bing Liu2, Yan Zhang3
1Tianjin Key Laboratory of Food Science and Health, School of Medicine, Nankai University, Tianjin, 300071, China; State Key Laboratory of Food Nutrition and Safety, Key Laboratory of Food Nutrition and Safety, Ministry of Education of China, Tianjin University of Science and Technology, Tianjin, 300457, China; ARC Centre of Excellence in Nanoscale Biophotonics (CNBP), Macquarie University, North Ryde, 2109, Australia.
Abstract:
In order to develop a sensitive immunoassay for Aflatoxin B1 (AFB1) monitoring, a hybridoma secreting anti-AFB1 monoclonal antibody with high binding affinity was screened. A new type of CdTe/CdS/ZnS quantum dot was synthesized and conjugated with an artificial antigen for use as a fluorescent probe in a simple one-step fluorescence immunoassay (FLISA). The developed FLISA allowed a sensitive determination of AFB1 in cereal samples in a wide linear range, from 0.08 to 1.97 ng mL-1, with a detection limit of 0.01 ng mL-1 in cereal samples. The corresponding immunoglobulin genes of the Fab fragment were cloned and sequenced, and expression of Fab was successfully verified in HEK293 cells, with a KD value of 1.09 × 10-7 mol L-1 for AFB1. To investigate the interactions between the antibody and AFB1, molecular docking, molecular dynamic simulation, and quantum-chemical computation were performed on AFB1 and a homology model of the Fab fragment. Our results showed that residues Ser32, Trp93, and Trp98 played the most important roles in the binding through hydrogen bonds formation, Pi-Pi stacked/Pi-alkyl interactions, and van der Waals interactions. In addition, the electrostatic potential study of AFB1 demonstrated that electrostatic interactions also played an important role in the recognition process. Results from theoretical studies provide guidance for hapten design and antibody improvement through genetic engineering.
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