Purification of the human fibroblast growth factor 2 using novel animal-component free materials.

Autor: Bolten SN; Institute of Technical Chemistry, Leibniz University of Hannover, Callinstraße 5, Hannover 30167, Germany., Knoll AS; Institute of Technical Chemistry, Leibniz University of Hannover, Callinstraße 5, Hannover 30167, Germany., Li Z; Institute of Technical Chemistry, Leibniz University of Hannover, Callinstraße 5, Hannover 30167, Germany., Gellermann P; Institute of Technical Chemistry, Leibniz University of Hannover, Callinstraße 5, Hannover 30167, Germany., Pepelanova I; Institute of Technical Chemistry, Leibniz University of Hannover, Callinstraße 5, Hannover 30167, Germany., Rinas U; Institute of Technical Chemistry, Leibniz University of Hannover, Callinstraße 5, Hannover 30167, Germany; Helmholtz Centre for Infection Research, Inhoffenstraße 7, Braunschweig 38124, Germany., Scheper T; Institute of Technical Chemistry, Leibniz University of Hannover, Callinstraße 5, Hannover 30167, Germany. Electronic address: scheper@iftc.uni-hannover.de.
Jazyk: angličtina
Zdroj: Journal of chromatography. A [J Chromatogr A] 2020 Aug 30; Vol. 1626, pp. 461367. Date of Electronic Publication: 2020 Jun 23.
DOI: 10.1016/j.chroma.2020.461367
Abstrakt: This paper analyzes the use of animal-component free chromatographic materials for the efficient purification of the human fibroblast growth factor 2 (hFGF-2). hFGF-2 is produced in Escherichia coli and purified via three different chromatography steps, which include a strong cation exchange chromatography as a capture step, followed by heparin affinity chromatography and an anion exchange chromatography as a polishing step. The affinity chromatography step is based on the animal-derived material heparin. Chemically produced ligands provide a viable alternative to animal-derived components in production processes, since they are characterized by a defined structure which leads to reproducible results and a broad range of applications. The alternative ligands can be assigned to adsorber of the mixed-mode chromatography (MMC) and pseudo-affinity chromatography. Eight different animal-component free materials used as adsorbers in MMC or pseudo-affinity chromatography were tested as a substitute for heparin. The MMCs were cation exchangers characterized with further functional residues. The ligands of the pseudo-affinity chromatography were heparin-like ligands which are based on heparin's molecular structure. The alternative methods were tested as a capture step and in combination with another chromatographic step in the purification procedure of hFGF-2. In each downstream step purity, recovery and yield were analysed and compared to the conventional downstream process. Two types of MMC - the column Foresight TM Nuvia TM cPrime TM from Bio-Rad Laboratories and the column HiTrap TM Capto TM MMC from GE Healthcare Life Sciences - can be regarded as effective animal-component free alternatives to the heparin - based adsorber.
Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2020. Published by Elsevier B.V.)
Databáze: MEDLINE
Externí odkaz: