Hansen M B, Lihme A, Spitali M, King D
UpFront Chromatography A/S, Lersø, Copenhagen, Denmark.
Bioseparation. 1999;8(1-5):189-93.
A novel group of mixed mode adsorbents has been developed for purification of monoclonal and polyclonal antibodies from a broad range of raw materials such as hybridoma cell culture, ascites fluid, animal sera, milk, whey and egg yolk. The aim of this study was to determine whether such mixed mode adsorbents were also useful for the recovery of recombinant proteins from microbial feedstocks. This paper describes the performance of one of these adsorbents for expanded bed capture of a human Fab fragment from recombinant E. Coli cell extracts. It is concluded that the mixed mode adsorbent binds the Fab fragment efficiently from crude extracts without any requirement for preconditioning the extract by for example de-salting or dilution. The capacity of the mixed mode adsorbent is approx. 12 mg Fab/ml matrix. The novel mixed mode adsorbent can be useful during production of highly purified Fab fragments as the first step in a purification scheme. In this respect the mixed mode adsorbent is advantageous over alternative commercially available ion-exchange materials which require pre-conditioning of cell extract for Fab' capture. Together with the concentration and clarification effect a significant enrichment of the Fab fragment is obtained in one single high yield operation.
已开发出一组新型混合模式吸附剂，用于从多种原料（如杂交瘤细胞培养物、腹水、动物血清、牛奶、乳清和蛋黄）中纯化单克隆抗体和多克隆抗体。本研究的目的是确定此类混合模式吸附剂是否也可用于从微生物原料中回收重组蛋白。本文描述了其中一种吸附剂用于从重组大肠杆菌细胞提取物中扩展床捕获人Fab片段的性能。得出的结论是，混合模式吸附剂可有效地从粗提取物中结合Fab片段，而无需对提取物进行例如脱盐或稀释等预处理。混合模式吸附剂的容量约为12 mg Fab/ml基质。这种新型混合模式吸附剂在生产高度纯化的Fab片段时作为纯化方案的第一步可能会很有用。在这方面，混合模式吸附剂优于其他市售离子交换材料，后者在捕获Fab'时需要对细胞提取物进行预处理。连同浓缩和澄清效果，在一次高产率操作中可实现Fab片段的显著富集。
