Coupling purification and on-column PEGylation of tumor necrosis factor alpha analogue

Trends in preparation of PEGylated protein drugs strive for simple, fast, and cheap processes, resulting in well-defined homogeneous products. We investigated the on-column PEGylation of tumor necrosis factor alpha (TNF-α), where purification and conjugation were performed in one step by using immob...

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Bibliographic Details
Published in:Analytical biochemistry 2012-11, Vol.430 (2), p.105-107
Main Authors: Milunović, Tatjana, Kunstelj, Menči, Fidler, Katarina, Anderluh, Gregor, Gaberc Porekar, Vladka
Format: Article
Language:English
Subjects:
affinity chromatography
chemical bonding
Chromatography, Affinity
drugs
Escherichia coli
Escherichia coli - metabolism
Escherichia coli Proteins - chemistry
IMAC
Immobilized Proteins - chemistry
Immobilized Proteins - isolation & purification
Immobilized Proteins - metabolism
On-column PEGylation
PEGylation
Polyethylene glycol
Polyethylene Glycols - chemistry
proteins
Recombinant Proteins - genetics
Recombinant Proteins - isolation & purification
Recombinant Proteins - metabolism
Tumor necrosis factor alpha
Tumor Necrosis Factor-alpha - chemistry
Tumor Necrosis Factor-alpha - isolation & purification
Tumor Necrosis Factor-alpha - metabolism
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Summary:Trends in preparation of PEGylated protein drugs strive for simple, fast, and cheap processes, resulting in well-defined homogeneous products. We investigated the on-column PEGylation of tumor necrosis factor alpha (TNF-α), where purification and conjugation were performed in one step by using immobilized metal affinity chromatography (IMAC). The same quality of the PEGylated product was obtained by the on-column approach starting from either the crude Escherichia coli protein extract or the purified protein. In comparison with the PEGylation in solution, the on-column approach resulted in more homogeneous PEGylated product. The on-column PEGylation reduces the number of production steps, costs, and preparation time.
ISSN:0003-2697
1096-0309
DOI:10.1016/j.ab.2012.08.013