Production and comprehensive quality control of recombinant human Interleukin-1β:: A case study for a process development strategy

被引：23

作者：

Block, Helena ^{[1
]}

Kubicek, Jan ^{[1
]}

Labahn, Joerg ^{[2
]}

Roth, Udo ^{[1
]}

Schaefer, Frank ^{[1
]}

机构：

[1] QIAGEN GmbH, D-40724 Hilden, Germany

[2] Res Ctr Julich, Inst Biol Struct, IBI 2, D-52425 Julich, Germany

来源：

PROTEIN EXPRESSION AND PURIFICATION | 2008年 / 57卷 / 02期

关键词：

Interleukin-1 beta (IL-1 beta); expression screening; protein production; X-ray crystallography; TAGZyme; DAPase; His-Tag cleavage; Immobilized-Metal Affinity Chromatography (IMAC); Ni-NTA;

D O I：

10.1016/j.pep.2007.09.019

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

We describe an efficient strategy to produce high-quality proteins by using a single large IMAC chromatography column and enzymatic His-tag removal via the TAGZyme system in pilot scale. Numerous quality assays demonstrated a high purity of the final product, the human cytokine Interleukin-1beta (IL-1 beta). The protein preparation was apparently free of host cell proteins, endotoxins, protease, and aggregates. The N-terminal amino acid sequence of IL-1 beta was in full agreement with the natural mature form of IL-1 beta. The homogeneity of the product was further shown by X-ray structure determination which confirmed the previously solved structure of the protein. We propose the applied workflow as a strategy for industrial production of protein-based biopharmaceuticals. (C) 2007 Elsevier Inc. All rights reserved.

引用

页码：244 / 254

页数：11

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