Preparation of mixed-mode adsorbent and its application for removing Protein A from antibody

doi:10.11949/j.issn.0438-1157.20171564

Abstract

Abstract:

A new mixed-mode chromatography (MMC) adsorbent, named Adhere NUPharose FF with functional ligand N-benzyl-N-merhylethanolamine was prepared and used for removing the leaked Protein A ligand while purifying antibody. The ligand presents hydrophobic, electrostatic and hydrogen bond interactions between Adhere NUPharose FF and adsorbate. The agarose beads were activated by allyl glycidyl ether and conditions of the activation and processes of ligand coupling on the agarose beads were optimized, leading to an activated density of 260 μmol·ml^-1 and a ligand density of 120 μmol·ml^-1. Meantime, the mixture of antibody and Protein A was used to test the ability of the prepared adsorbent to remove Protein A from antibody. The data showed that it could remove 83.1% Protein A from antibody preparation, and supply feasible method to remove Protein A from antibody product.

Key words: mixed-mode chromatography, preparation, antibody, Protein A, ligand density

摘要：

针对抗体纯化过程中脱落蛋白A的清除，设计和制备了以N-苄基-N-甲基乙醇胺为功能配基的混合模式层析介质Adhere NUPharose FF，该配基兼有疏水、静电和氢键作用。采用烯丙基缩水甘油醚活化法，并对琼脂糖微球基质的活化和配基偶联条件进行了优化，优化后活化密度达260 μmol·ml^-1以上，配基密度达120 μmol·ml^-1。同时以抗体和蛋白A的混合物为研究对象来考察该介质对抗体中蛋白A的清除能力，结果表明对蛋白A的有效去除率达到83.1%，为抗体制品中蛋白A的清除提供了一种可行性方法。

关键词: 混合模式介质, 制备, 抗体, 蛋白A, 配基密度

CLC Number:

TQ028.8

YAN Jun, QIAN Yongchang, YAO Shanjing, LIN Dongqiang. Preparation of mixed-mode adsorbent and its application for removing Protein A from antibody[J]. CIESC Journal, 2018, 69(7): 3190-3197.

严军, 钱永常, 姚善泾, 林东强. 混合模式介质的制备及用于清除抗体中蛋白A[J]. 化工学报, 2018, 69(7): 3190-3197.

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