动脉采血,4℃ 过夜后,离心取上清,即为抗血清。经过盐析,Protein A/G亲和层析,非IgG类蛋白流穿,IgG通过Fc端结合到Protein A结构域上,过量的结合缓冲液,平衡后低pH洗脱,有尖锐UV洗脱峰。收集洗脱液,并平衡至中性pH。 经过缓冲液置换,浓缩,Nanodrop One C UV以及Bradford法定量,并标记生物素或者辣根过氧化物酶,用于HCP的结合验证实验。
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