Journal Article

PreJuSER-5173

http://join2-wiki.gsi.de/foswiki/pub/Main/Artwork/join2_logo100x88.png Production and comprehensive quality control of recombinant human interleukin-1 beta: A case study for a process development strategy

Block, H. ; Kubicek, J. ; Labahn, J.FZJ* ; Roth, U. ; Schaefer, F.

2008
Academic Press Orlando, Fla.

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Please use a persistent id in citations: doi:10.1016/j.pep.2007.09.019

Abstract: 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-1beta). The protein preparation was apparently free of host cell proteins, endotoxins, protease, and aggregates. The N-terminal amino acid sequence of IL-1beta was in full agreement with the natural mature form of IL-1beta. 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.

Keyword(s): Amino Acid Sequence (MeSH) ; Biotechnology: methods (MeSH) ; Chromatography, Affinity (MeSH) ; Electrophoresis, Gel, Two-Dimensional (MeSH) ; Endodeoxyribonucleases: isolation & purification (MeSH) ; Endoribonucleases: isolation & purification (MeSH) ; Endotoxins: isolation & purification (MeSH) ; Exopeptidases: isolation & purification (MeSH) ; Histidine: metabolism (MeSH) ; Humans (MeSH) ; Interleukin-1beta: biosynthesis (MeSH) ; Interleukin-1beta: chemistry (MeSH) ; Interleukin-1beta: isolation & purification (MeSH) ; Models, Molecular (MeSH) ; Molecular Sequence Data (MeSH) ; Oligopeptides: metabolism (MeSH) ; Recombinant Proteins: biosynthesis (MeSH) ; Recombinant Proteins: chemistry (MeSH) ; Recombinant Proteins: isolation & purification (MeSH) ; Sequence Analysis, Protein (MeSH) ; Endotoxins ; His-His-His-His-His-His ; Interleukin-1beta ; Oligopeptides ; Recombinant Proteins ; Histidine ; Endodeoxyribonucleases ; Endoribonucleases ; Serratia marcescens nuclease ; Exopeptidases ; J ; Interleukin-1 beta (IL-1 beta) (auto) ; expression screening (auto) ; protein production (auto) ; X-ray crystallography (auto) ; TAGZyme (auto) ; DAPase (auto) ; His-Tag cleavage (auto) ; Immobilized-Metal Affinity Chromatography (IMAC) (auto) ; Ni-NTA (auto)

Note: Record converted from VDB: 12.11.2012

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Contributing Institute(s):

1. Molekulare Biophysik (ISB-2)

Research Program(s):

1. Programm Biosoft (N03)

Appears in the scientific report 2009
Notes: übergangsweise lautet die FE-Bezeichnung für das ISB-2 N03

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