单克隆抗体TNT-1的纯化、F(ab)2片段化及其免疫活性的测定
TNT-1是-种鼠IgG2a单抗,已用于肿瘤病人的临床试验。本文将介绍从腹水中进一步纯化TNT-1的方法及制备其F(ah′)2片段的条件。经Protein A亲和柱层析从小鼠腹水中提取出的lgG,再通过阳离子交换树脂柱Mono S,用缓冲液A(20 mmol/L MES,pH6.3)和缓冲液B(lmol/L NaCI),在快速蛋白液相色层(FPLC)系统上进行梯度洗脱分离,可制得纯净的TNT-1抗体,其免疫活性可达80%~85%;经纯化的TNT-1,用胃蛋白酶消化,制备F(ah′)2片段,其消化条件是 ...
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