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首页> 外文期刊>Analytical Letters >Immobilization of Chymotrypsin on Silica Beads Based on High Affinity and Specificity Aptamer and Its Applications
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Immobilization of Chymotrypsin on Silica Beads Based on High Affinity and Specificity Aptamer and Its Applications

机译：基于高亲和力和适体的胰凝乳蛋白酶固定化硅胶微珠及其应用

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摘要

Aptamers with high affinity and specificity to targets, bring new approaches to immobilizing proteins or enzymes. In this work, a group of single-stranded DNA aptamers specific for chymotrypsin were obtained by SELEX method in vitro. After investigation and characterization of all aptamers, AptC.1 (abbreviation for the aptamer with the highest affinity for chymotrypsin) was selected and grafted onto silica matrix with the help of glutaraldehyde as linker, and used subsequently to immobilize chymotrypsin. Specifically, it is shown in experiment that, 12.65 μg of chymotrypsin could be immobilized on 10 mg of AptC.1-Silica in 10 mM pH 8.0 borate solutions, and the activity of immobilized enzyme was not inhibited. Bovine serum albumin, myoglobin and cytochrome c were introduced to investigate the enzymatic performance of prepared immobilized chymotrypsin reactor. All these results demonstrated that aptamer could serve as a potential medium for the immobilization of proteins or enzymes.
机译：对靶标具有高亲和力和特异性的适体，带来了固定蛋白质或酶的新方法。在这项工作中，通过SELEX方法体外获得了一组针对胰凝乳蛋白酶的单链DNA适体。在对所有适体进行研究和鉴定后，选择AptC.1（对胰凝乳蛋白酶具有最高亲和力的适体的缩写），并在戊二醛的帮助下将其嫁接到硅胶基质上，随后用于固定胰凝乳蛋白酶。具体地，在实验中表明，在10mM pH 8.0硼酸盐溶液中，可以将12.65μg的胰凝乳蛋白酶固定在10mg的AptC.1-二氧化硅上，并且不抑制固定的酶的活性。介绍了牛血清白蛋白，肌红蛋白和细胞色素c，以研究制备的固定化胰凝乳蛋白酶反应器的酶性能。所有这些结果表明，适体可以作为固定蛋白质或酶的潜在介质。

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