Renewable enzyme reactors based on beds of artificial gel antibodies

Maria Hjertén , Melinda Rezeli , Ferenc Kilár , Stellan Hjertén
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引用次数: 3

Abstract

A novel approach is described for the synthesis of beds for enzyme reactors. The method is based on the use of artificial antibodies in the form of polyacrylamide gel particles with diameters around 0.1–0.3 mm. These gel particles mimic protein antibodies, raised in experimental animals, in the sense that they selectively recognize and adsorb only the protein present during the preparation of the “antibodies”. The gel antibodies have several advantages over conventional protein antibodies, which can be taken advantage of in the design of enzyme reactors; for instance, if upon prolonged use the immobilized enzyme loses its activity it can easily be replaced by an active enzyme, which is not possible when the enzyme is immobilized via a conventional protein antibody (a new bed with immobilized protein antibodies must be prepared); and equally or more remarkable: the enzyme can be applied in the form of a non-purified extract since the selectivity of the artificial gel antibodies is so high that they will “fish-out” the enzyme, but no other proteins in the extract. In addition, no preconcentration of the enzyme solution is required prior to the immobilization, since the enzyme is enriched at the top of the column upon the application. These unique properties make enzyme reactors based on artificial gel antibodies very attractive, also in process chromatography. The potential application range of the artificial gel antibodies is enormous since the same method for their synthesis can be used independent of the structure and the size of the “antigen”; for instance, renewable biosensors based on gel antibodies for the selective detection of protein biomarkers, as well as pathogenic viruses, bacteria, and spores (for instance Anthrax) should not be difficult to design.

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基于人工凝胶抗体床的可再生酶反应器
介绍了一种合成酶反应器床的新方法。该方法基于使用人工抗体,其形式为聚丙烯酰胺凝胶颗粒,直径约为0.1-0.3毫米。这些凝胶颗粒模仿在实验动物中培养的蛋白质抗体,在某种意义上,它们选择性地识别和吸附“抗体”制备过程中存在的蛋白质。与传统的蛋白质抗体相比,凝胶抗体具有许多优点,可用于酶反应器的设计;例如,如果在长时间使用后,固定化酶失去了活性,它可以很容易地被活性酶取代,这是不可能的,当酶通过传统的蛋白质抗体固定化(必须准备一个新的床固定化蛋白质抗体);同样或更值得注意的是:这种酶可以以非纯化提取物的形式应用,因为人工凝胶抗体的选择性非常高,它们会“捞出”这种酶,但在提取物中没有其他蛋白质。此外,在固定化之前不需要预先浓缩酶溶液,因为酶在应用时在柱的顶部富集。这些独特的性质使得基于人工凝胶抗体的酶反应器非常有吸引力,在过程色谱中也是如此。人工凝胶抗体的潜在应用范围是巨大的,因为它们的合成方法可以独立于“抗原”的结构和大小;例如,基于凝胶抗体的可再生生物传感器,用于选择性检测蛋白质生物标志物,以及致病性病毒、细菌和孢子(例如炭疽),设计起来应该不难。
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