Production and comprehensive quality control of recombinant human Interleukin-1beta: a case study for a process development strategy.
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.
Block
Helena
H
QIAGEN GmbH, Qiagen Strasse 1, 40724 Hilden, Germany.
Kubicek
Jan
J
Labahn
Jörg
J
Roth
Udo
U
Schäfer
Frank
F
D000595:Amino Acid Sequence; D001709:Biotechnology / Q000379:methods*; D002846:Chromatography, Affinity; D015180:Electrophoresis, Gel, Two-Dimensional; D004706:Endodeoxyribonucleases / Q000302:isolation & purification; D004722:Endoribonucleases / Q000302:isolation & purification; D004731:Endotoxins / Q000302:isolation & purification; D020689:Exopeptidases / Q000302:isolation & purification; D006639:Histidine / Q000378:metabolism; D006801:Humans; D053583:Interleukin-1beta / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D020539:Sequence Analysis, Protein
D004731:Endotoxins; C471213:His-His-His-His-His-His; D053583:Interleukin-1beta; D009842:Oligopeptides; D011994:Recombinant Proteins; D006639:Histidine; D004706:Endodeoxyribonucleases; D004722:Endoribonucleases; C021835:Serratia marcescens nuclease; D020689:Exopeptidases