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

Research output: Contribution to journalArticleResearchpeer review

Authors

  • Svenja Nicolin Bolten
  • Anne Sophie Knoll
  • Zhaopeng Li
  • Pia Gellermann
  • Iliyana Pepelanova
  • Ursula Rinas
  • Thomas Scheper

Research Organisations

External Research Organisations

  • Helmholtz Centre for Infection Research (HZI)
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Details

Original languageEnglish
Article number461367
JournalJournal of Chromatography A
Volume1626
Early online date23 Jun 2020
Publication statusPublished - 30 Aug 2020

Abstract

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 ForesightTM NuviaTM cPrimeTM from Bio-Rad Laboratories and the column HiTrapTM CaptoTM MMC from GE Healthcare Life Sciences - can be regarded as effective animal-component free alternatives to the heparin - based adsorber.

Keywords

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

ASJC Scopus subject areas

Sustainable Development Goals

Cite this

Purification of the human fibroblast growth factor 2 using novel animal-component free materials. / Bolten, Svenja Nicolin; Knoll, Anne Sophie; Li, Zhaopeng et al.
In: Journal of Chromatography A, Vol. 1626, 461367, 30.08.2020.

Research output: Contribution to journalArticleResearchpeer review

Bolten SN, Knoll AS, Li Z, Gellermann P, Pepelanova I, Rinas U et al. Purification of the human fibroblast growth factor 2 using novel animal-component free materials. Journal of Chromatography A. 2020 Aug 30;1626:461367. Epub 2020 Jun 23. doi: 10.1016/j.chroma.2020.461367
Bolten, Svenja Nicolin ; Knoll, Anne Sophie ; Li, Zhaopeng et al. / Purification of the human fibroblast growth factor 2 using novel animal-component free materials. In: Journal of Chromatography A. 2020 ; Vol. 1626.
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abstract = "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 ForesightTM NuviaTM cPrimeTM from Bio-Rad Laboratories and the column HiTrapTM CaptoTM MMC from GE Healthcare Life Sciences - can be regarded as effective animal-component free alternatives to the heparin - based adsorber.",
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AU - Pepelanova, Iliyana

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