Department of Protein Biochemistry, Roche Research Center, Hoffmann-La Roche, Inc., Nutley, New Jersey 07110, USA.
Biotechnol Bioeng. 1992 Aug;40(5):564-71. doi: 10.1002/bit.260400503.
A systematic approach to the design and development of membrane-based immunoaffinity systems for the purification of recombinant proteins is presented. The preparation and characterization of immunoaffinity membranes are described. The immunoaffinity purification process for recombinant interferon-alpha2a is used as a model system to determine the operational parameters in membrane-based immunoaffinity chromatography. The high volumetric throughput of membranes, combined with the typically fastbinding kinetics of antigen-antibody interactions, enable the purification of recombinant proteins from dilute feed stream in less time, using less antibody than conventional systems. Three recombinant proteins, human interferon-alpha2a, interleukin-2, and interleukin-2 receptor, have been purified efficiently employing membrane-based immunoaffinity chromatography. Overall, membrane-based immunoaffinity chromatography is shown to be a viable and scalable method, ideal for the industrial-scale production of recombinant proteins.
提出了一种用于膜基免疫亲和系统设计和开发的系统方法，用于重组蛋白的纯化。描述了免疫亲和膜的制备和表征。以重组干扰素-α2a 的免疫亲和纯化过程为例，确定了膜基免疫亲和层析中的操作参数。膜的高体积通量，加上抗原 - 抗体相互作用的典型快速结合动力学，使得能够在更短的时间内从稀释的进料流中纯化重组蛋白，使用的抗体比传统系统少。使用基于膜的免疫亲和层析有效地纯化了三种重组蛋白，人干扰素-α2a、白细胞介素-2 和白细胞介素-2 受体。总的来说，膜基免疫亲和层析是一种可行且可扩展的方法，非常适合重组蛋白的工业规模生产。
