期刊
APPLIED SURFACE SCIENCE
卷 355, 期 -, 页码 570-576出版社
ELSEVIER
DOI: 10.1016/j.apsusc.2015.05.020
关键词
Dextran hydrogel surface; Small molecule detection; Surface plasmon resonance imaging; Protein array
类别
资金
- National Major Scientific Instruments and Equipments Development Project [2011YQ03012405]
- National Natural Science Foundation of China [61077064/60921001]
- Program of International ST Cooperation [2010DFB33880]
The label-free and sensitive detection of small molecule drugs on SPRi is still a challenging task, mainly due to the limited surface immobilization capacity of the sensor. In this research, a dextran hydrogel-coated gold sensor chip for SPRi was successfully fabricated via photo-cross-linking for enhanced surface immobilization capacity. The density of the dextran hydrogel was optimized for protein immobilization and sensitive small molecule detection. The protein immobilization capacity of the hydrogel was 10 times greater than a bare gold surface, and 20 times greater than an 11-mercaptoundecanoic acid (MUA) surface. Such a drastic improvement in immobilization capacity allowed the SPRi sensor to detect adequate response signals when probing small molecule binding events. The binding signal of 4 nM liquid-phase biotin to streptavidin immobilized on the dextran surface reached 435 RU, while no response was observed on bare gold or MUA surfaces. The dextran hydrogel-coated SPRi sensor was also applied in a kinetic study of the binding between an immunosuppressive drug (FK506) and its target protein (FKBP12) in a high-throughput microarray format. The measured binding affinity was shown to be consistent with reported literature values, and a detection limit of 0.5 nM was achieved. (c) 2015 Elsevier B.V. All rights reserved.
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