Coupling purification and on-column PEGylation of tumor necrosis factor alpha analogue |
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| Authors: | Tatjana Milunović Menči Kunstelj Katarina Fidler Gregor Anderluh Vladka Gaberc Porekar |
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| Affiliation: | 1. Laboratory for Molecular Biology and Nanobiotechnology, National Institute of Chemistry, 1000 Ljubljana, Slovenia;2. Sandoz Biopharmaceuticals–Lek Pharmaceuticals d.d., SI-1234 Menge?, Slovenia;3. Department of Biology, Biotechnical Faculty, University of Ljubljana, 1000 Ljubljana, Slovenia |
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| Abstract: | ![]()
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. |
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