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温控黏附碳水化合物功能化微凝胶膜:大肠杆菌和凝集素结合研究。

Temperature-Controlled Adhesion to Carbohydrate Functionalized Microgel Films: An E. coli and Lectin Binding Study.

机构信息

Institute of Organic and Macromolecular Chemistry, Heinrich-Heine-University Düsseldorf, Universitätsstraße 1, Düsseldorf, 40225, Germany.

出版信息

Macromol Biosci. 2021 Apr;21(4):e2000386. doi: 10.1002/mabi.202000386. Epub 2021 Feb 19.

Abstract

The preparation of thermoresponsive mannose functionalized monolayers of poly(N-isopropylacrylamide) microgels and the analysis of the specific binding of concanavalin A (ConA) and E. coli above and below the lower critical solution temperature (LCST) are shown. Via inhibition and direct binding assays it is found that ConA binding is time-dependent, where at short incubation times binding is stronger above the LCST. Given larger incubation times, the interaction of ConA to the microgel network is increased below the LCST when compared to temperatures above the LCST, possibly due to increased ConA diffusion and multivalent binding in the more open microgel network below the LCST. For E. coli, which presents only monovalent lectins and is too large to diffuse into the network, binding is always enhanced above the LCST. This is due to the larger mannose density of the microgel layer above the LCST increasing the interaction to E. coli. Once bound to the microgel layer above the LCST, E. coli cannot be released by cooling down below the LCST. Overall, this suggests that the carbohydrate presenting microgel layers enable specific binding where the temperature-induced transition between swollen and collapsed microgels may increase or decrease binding depending on the receptor size.

摘要

本文展示了聚(N-异丙基丙烯酰胺)微凝胶的温敏性甘露糖功能化单层的制备,以及在低临界溶液温度(LCST)之上和之下分析伴刀豆球蛋白 A(ConA)和大肠杆菌的特异性结合。通过抑制和直接结合实验发现,ConA 结合具有时间依赖性,在 LCST 之上短孵育时间内结合更强。考虑到更长的孵育时间,与 LCST 之上的温度相比,ConA 与微凝胶网络的相互作用在 LCST 之下增加,这可能是由于在 LCST 之下更开放的微凝胶网络中 ConA 扩散和多价结合增加。对于仅呈现单价凝集素且太大而无法扩散到网络中的大肠杆菌,结合总是在 LCST 之上增强。这是由于 LCST 之上微凝胶层的甘露糖密度较大,增加了与大肠杆菌的相互作用。一旦与 LCST 之上的微凝胶层结合,大肠杆菌就不能通过冷却到 LCST 以下而释放。总体而言,这表明糖展示微凝胶层能够实现特异性结合,其中溶胀和塌陷微凝胶之间的温度诱导转变可能会增加或减少结合,具体取决于受体的大小。

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