Naslov
Purification of human tumour necrosis factor by membrane chromatography

Autori
Luksa, Janja ; Menart, Viktor ; Milicic, Stjepan ; Kus, Breda ; Gaberc-Porekar, Vladimira ; Josic, Djuro

Izvornik
Journal of Chromatography A (0021-9673) 661 (1994), 1-2; 161-168

Vrsta, podvrsta i kategorija rada
Radovi u časopisima, članak, znanstveni

Ključne riječi
tumour necrosis factor ; membrane chromatography

Sažetak
The recombinant human tumour necrosis factor α from an extract of Escherichia coli was enriched to homogeneity according to specific activity and sodium dodecyl sulphate-polyacrylamide gel electrophoresis by purification using anion-exchange HPLC and hydrophobic interaction HPLC. Parallel experiments with the same separation methods, but carried out with membrane chromatography on compact discs, gave similar results in terms of yield and purity of the product. The active form of the protein is a trimer. The second isolation step, hydrophobic interaction chromatography, causes dissociation of the trimer into monomers and a partial loss of the biological activity of the protein. The phenomenon occurs on both the column and the disc. This in turn indicates strongly that the dissociation of the protein is a consequence of interaction between the sample and the hydrophobic ligand, and is not caused by non-specific interaction with the matrix.

Izvorni jezik
Engleski

Znanstvena područja
Biotehnologija u biomedicini (prirodno područje, biomedicina i zdravstvo, biotehničko područje)

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