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目的 从猪脑中提取纯化谷氨酸脱羧酶（ Ｇ Ａ Ｄ） ，用于１ 型糖尿病患者 Ｇ Ａ Ｄ 抗体的测定。方法 以猪脑制备 Ｇ Ａ Ｄ 粗酶，经 Ｓｅｐｈａｄｅｘ Ｇ１００ 、 Ｄ Ｅ Ａ Ｅ Ｓｅｐｈａｃｅｌ 和羟基磷灰石凝胶层析分离纯化，结合十二烷基硫酸钠聚丙烯酰胺凝胶电泳（ Ｓ Ｄ Ｓ Ｐ Ａ Ｇ Ｅ） 测定相对分子质量、同位素法测定 Ｇ Ａ Ｄ酶活性，建立了 Ｇ Ａ Ｄ 纯化方法。结果 Ｇ Ａ Ｄ 粗酶经纯化后的单位蛋白量的酶活性增加了１１８ 倍。经 Ｓ Ｄ Ｓ Ｐ Ａ Ｇ Ｅ 分析，为一条相对分子质量约６４ ０００ 的蛋白条带。结论 用多步层析分离法从猪脑中提取纯化 Ｇ Ａ Ｄ 是一种简便可行的方法。
Objective To extract purified glutamic acid decarboxylase (G A D) from porcine brain for the determination of G A D antibody in type 1 diabetic patients. Methods G A D crude enzyme was prepared from porcine brain and purified by Sephadex G100, D E A Sephacel and hydroxyapatite gel chromatography. SDS-PAGE coupled with sodium dodecyl sulfate-polyacrylamide gel electrophoresis S P A G E) determination of relative molecular mass, G A D enzyme activity was determined by isotope method, G A D purification method was established. As a result, the amount of purified protein per unit amount of GDA crude enzyme increased by 118-fold. After S D S P A G E analysis, a relative molecular mass of about 64 000 protein bands. Conclusion The purification of G A D from porcine brain by multi-step chromatography is a simple and feasible method.