基因工程药物中过敏性杂质青霉噻唑蛋白的评价体系

EVALUATION OF PENICILLOYL PROTEINS OF ALLERGIC IMPURITY IN GENE ENGINEERING DRUGS

对基因工程药品中过敏性杂质青霉噻唑蛋白（ｐｅｎｉｃｉｌｏｙｌｐｒｏｔｅｉｎ）的评价体系进行了研究。将酶联免疫吸附法（ｅｎｚｙｍｅｌｉｎｋｅｄｉｍｍｕｎｏｓｏｒｂｅｎｔａｓａｙ，ＥＬＩＳＡ）和被动皮肤过敏（ｐａｓｉｖｅｃｕｔａｎｅｏｕｓａｎａｐｈｙｌａｘｉｓ，ＰＣＡ）实验联用，前者具有较高的灵敏度和特异性，对样品中结合青霉素的绝对检出量小于０３ｐｐｍ；后者为整体动物实验，是经典的Ｉ型过敏反应动物模型；从体内、外两方面构成了一个较为完整的评价体系。通过对近年来国内报批的新药包括促红细胞生成素（ＥＰＯ）、人粒细胞集落刺激因子（ＧＣＳＦ）、人粒细胞巨噬细胞集落刺激因子（ＧＭＣＳＦ）、新型人白细胞介素２（１２５ＳｅｒＩＬ２）等基因工程药品的分析评价。

It is well known that penicillins are necessary for prevention of contamination during cell culture or fermentation in the production of gene engineering drugs. As penicillins are easily combined with proteins forming allergic impurities, the benzyl penicilloyl proteins in aqueous solution, an evaluation system should be established for control of the gene engineering drugs. An enzyme linked immunosorbent assay (ELISA) with high specificity and high sensitivity in vitro, by which the conjugated penicillins of 03 ppm in a sample could be detected and a passive cutaneous anaphylaxis (PCA) test, a classical method for evaluation of allergic reaction in vivo are integral parts of this system.Some domestic gene engineering drugs such as erythropoietin (EPO), GCSF, GMCSF and 125SerIL2 are evaluated with this system. No matter whether the expression products was produced in secretion manner or in inclusion body, it was possible that some residues of allergic impurities may remain in the finished products if an unsuitable process of isolation and purification was used. The maximum absolute content of the conjugated penicillins in a sample was up to about one per thousand.