用单克隆抗体特异性固定化一种酶:固定化豚鼠肝脏转谷氨酰胺酶。

Journal of applied biochemistry Pub Date : 1984-08-01
K Ikura, K Okumura, M Yoshikawa, R Sasaki, H Chiba
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引用次数: 0

摘要

以豚鼠肝转谷氨酰胺酶为例,研究了一种简单的酶固定化方法。该方法包括两个步骤;通过琼脂糖凝胶球载体将单克隆抗体与酶结合而不抑制酶的活性,并通过抗原-抗体免疫反应将酶蛋白固定在偶联的单克隆抗体上。以下是这种方法的优点。(1)固定化后活性恢复非常高;(2)粗酶制剂中的酶蛋白可以特异性固定化;(3)通过尿素缓冲处理,固定化后的酶蛋白很容易被新鲜酶蛋白取代。
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Specific immobilization of an enzyme by monoclonal antibody: immobilization of guinea pig liver transglutaminase.

A simple method for enzyme immobilization using a monoclonal antibody raised against the enzyme was studied, taking guinea pig liver transglutaminase as an example. This method consists of two steps; a conjugation of monoclonal antibodies, which bind with the enzyme without inhibitory effect on the activity, with agarose gel bead supports, and immobilization of enzyme proteins onto the conjugated monoclonal antibodies through the antigen-antibody immunoreaction. The following are the advantages of this method. (i) Activity recovery after the immobilization was very high, (ii) enzyme proteins in crude enzyme preparation could be immobilized specifically, and (iii) immobilized enzymes could be replaced easily by fresh enzyme proteins through a urea-buffer treatment.

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