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一种用于构建多价表面的简便原位微流控方法:迈向功能糖组学。

A facile in situ microfluidic method for creating multivalent surfaces: toward functional glycomics.

机构信息

KIST Europe, Korea Institute of Science and Technology, Campus E7 1, 66123 Saarbruecken, Germany.

出版信息

Lab Chip. 2012 Apr 21;12(8):1500-7. doi: 10.1039/c2lc21217j. Epub 2012 Mar 9.

Abstract

An in situ method of modifying the chemistry and topology of microfluidic surfaces in order to mimic the cellular environment is described. The binding of functionalised microbeads to microfluidic channels allows the surface-to-volume ratio of the system, and thus the number of biomolecules available for reaction, to be vastly increased, thereby enhancing the sensitivity of biochemical analyses. The sensitivity and specificity of the technique were first investigated via the study of carbohydrate-protein interactions. Beads featuring hydrazide moieties were adhered to the channel surface, after which carbohydrates (galactose and mannose) were bound to the beads in situ and reacted with fluorescently labelled proteins. Results showed a six-fold increase in fluorescent signal compared to the same process performed on a glass surface without the presence of beads, thereby demonstrating the increase in valence afforded by the method. In a subsequent study, beads, modified with galactose moieties via the in situ functionalisation technique, were used to perform studies of colon tumour cells from a cell sample. Here, the carcinoma cells exhibited superior adhesion than the normal cells due to an increased expression of active galactose receptors, thereby demonstrating the success of the biofunctionalisation method for investigating cellular mechanisms.

摘要

本文描述了一种在原地改变微流控表面化学和拓扑结构以模拟细胞环境的方法。功能化微珠与微流控通道的结合允许系统的表面积与体积比(即可用于反应的生物分子数量)大大增加,从而提高生化分析的灵敏度。该技术的灵敏度和特异性首先通过碳水化合物-蛋白质相互作用的研究来研究。将带有酰肼部分的珠子附着在通道表面上,然后将碳水化合物(半乳糖和甘露糖)原位结合到珠子上,并与荧光标记的蛋白质反应。结果表明,与在没有珠子的情况下在玻璃表面上进行相同的过程相比,荧光信号增加了六倍,从而证明了该方法提供的配位数增加。在随后的一项研究中,通过原位功能化技术用半乳糖部分修饰的珠子用于从细胞样本中进行结肠癌肿瘤细胞的研究。在这里,由于活性半乳糖受体的表达增加,癌细胞的粘附性优于正常细胞,从而证明了生物功能化方法用于研究细胞机制的成功。

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