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

Animal-component free Heparin affinity chromatography Human fibroblast growth factor 2 (hFGF-2) Mixed-mode chromatography Pseudo-affinity chromatography

Journal

Journal of chromatography. A
ISSN: 1873-3778
Titre abrégé: J Chromatogr A
Pays: Netherlands
ID NLM: 9318488

Informations de publication

Date de publication:
30 Aug 2020
Historique:
received: 05 03 2020
revised: 11 06 2020
accepted: 22 06 2020
entrez: 17 8 2020
pubmed: 17 8 2020
medline: 22 9 2020
Statut: ppublish

Résumé

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

Identifiants

pubmed: 32797846
pii: S0021-9673(20)30644-0
doi: 10.1016/j.chroma.2020.461367
pii:
doi:

Substances chimiques

Ligands 0
Fibroblast Growth Factor 2 103107-01-3
Heparin 9005-49-6

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

461367

Informations de copyright

Copyright © 2020. Published by Elsevier B.V.

Déclaration de conflit d'intérêts

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.

Auteurs

Svenja Nicolin Bolten (SN)

Institute of Technical Chemistry, Leibniz University of Hannover, Callinstraße 5, Hannover 30167, Germany.

Anne-Sophie Knoll (AS)

Institute of Technical Chemistry, Leibniz University of Hannover, Callinstraße 5, Hannover 30167, Germany.

Zhaopeng Li (Z)

Institute of Technical Chemistry, Leibniz University of Hannover, Callinstraße 5, Hannover 30167, Germany.

Pia Gellermann (P)

Institute of Technical Chemistry, Leibniz University of Hannover, Callinstraße 5, Hannover 30167, Germany.

Iliyana Pepelanova (I)

Institute of Technical Chemistry, Leibniz University of Hannover, Callinstraße 5, Hannover 30167, Germany.

Ursula 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.

Thomas Scheper (T)

Institute of Technical Chemistry, Leibniz University of Hannover, Callinstraße 5, Hannover 30167, Germany. Electronic address: scheper@iftc.uni-hannover.de.

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Classifications MeSH