公开(公告)号：WO2013082518A1

公开(公告)日：2013-06-06

申请号：PCT/US2012/067427

申请日：2012-11-30

Applicant: LIFE TECHNOLOGIES CORPORATION

Inventor： JONES, Meredith, Bauman ,  VASU, Sanjay ,  JUDD, David

IPC: B01D15/38 ,  G01N30/14

CPC classification number: G01N30/04 ,  B01D15/12 ,  B01D15/20 ,  B01D15/3828 ,  G01N2030/8831

Abstract: Compositions and methods for increasing protein recovery and/ or reducing metal ion stripping in IMAC chromatography or from resins used therein are provided. Samples like spent/ conditioned cell culture media or cell extracts containing native or recombinant proteins, were supplemented with metal ions before loading on to IMAC resins or columns to reduce metal ion stripping and/ or to increase protein recovery. The compositions and methods discussed herein work for a variety of metal affinity resins. Use of the compositions and methods disclosed may result in reusability of columns or resins, and improve the yield, purity and cost-effectiveness of using metal-ion based chromatography. The process is scalable to large-scale commercial levels.

Abstract translation: 提供了用于增加IMAC色谱中或从其中使用的树脂中的蛋白质回收和/或还原金属离子剥离的组合物和方法。 含有天然或重组蛋白质的废弃/调节细胞培养基或细胞提取物的样品在加载到IMAC树脂或柱之前补充金属离子以减少金属离子剥离和/或增加蛋白质回收。 本文讨论的组合物和方法适用于各种金属亲和树脂。 使用所公开的组合物和方法可以导致柱或树脂的可再利用性，并且提高使用基于金属离子的色谱法的产率，纯度和成本效益。 该过程可扩展到大规模的商业级别。