Abstract
A novel approach for preparing carbohydrate chips based on polydopamine (PDA) surface to study carbohydrate–lectin interactions by quartz crystal microbalance (QCM) biosensor instrument has been developed. The amino-carbohydrates were immobilized on PDA-coated quartz crystals via Schiff base reaction and/or Michael addition reaction. The resulting carbohydrate-chips were applied to QCM biosensor instrument with flow-through system for real-time detection of lectin–carbohydrate interactions. A series of plant lectins, including wheat germ agglutinin (WGA), concanavalin A (Con A), Ulex europaeus agglutinin I (UEA-I), soybean agglutinin (SBA), and peanut agglutinin (PNA), were evaluated for the binding to different kinds of carbohydrate chips. Clearly, the results show that the predicted lectin selectively binds to the carbohydrates, which demonstrates the applicability of the approach. Furthermore, the kinetics of the interactions between Con A and mannose, WGA and N-Acetylglucosamine were studied, respectively. This study provides an efficient approach to preparing carbohydrate chips based on PDA for the lectin–carbohydrate interactions study.
Highlights
Carbohydrates located on the surfaces of cells play a crucial role in cell interactions, cell communication, cell proliferation, as well as cell death, where the interactions between carbohydrates and proteins are identified to be essential important in biological processes [1,2,3]
The quartz crystal microbalance (QCM) biosensor, which is based on the effect of piezoelectric, has turned out to be an efficient analytical tool for the assessment of carbohydrate–protein interactions with protein chips, carbohydrate chips, and cell chips [13,14,15,16,17,18,19]
Following PDA coating, the aminated carbohydrates were immobilized on the PDA coated chip surface via Schiff base and/or Michael addition reaction to yield the carbohydrate chips, respectively
Summary
Carbohydrates located on the surfaces of cells play a crucial role in cell interactions, cell communication, cell proliferation, as well as cell death, where the interactions between carbohydrates and proteins are identified to be essential important in biological processes [1,2,3]. Oscar et al [27,28] prepared a rapidly functionalized carbohydrate chip, which was the covalent immobilization of carbohydrate on the surface of polymer chips via click reaction and the PFPA azide photoligation. A protein chip on the surface of rapid functional chip using PDA was successfully prepared, which was used to analyze the kinetics of protein-protein binding process [35]. We have prepared carbohydrate chips based on PDA surface to evaluate carbohydrate–lectin interactions by QCM. Theaminated monosaccharide derivatives were covalently immobilized on the surface of PDA coated chip via Schiff base reaction and/or Michael addition reactions, respectively, to give carbohydrate chips, which were used for the study of interaction and kinetics of carbohydrate and protein. This research provides a novel way to preparing carbohydrate chips based on PDA for the study of lectin–carbohydrate interactions
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