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

authored by: Svenja Nicolin Bolten, Anne Sophie Knoll, Zhaopeng Li, Pia Gellermann, Iliyana Pepelanova, Ursula Rinas, Thomas Scheper
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 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.
Organisation(s): Institute of Technical Chemistry
External Organisation(s): Helmholtz Centre for Infection Research (HZI)
Type: Article
Journal: Journal of Chromatography A
Volume: 1626
ISSN: 0021-9673
Publication date: 30.08.2020
Publication status: Published
Peer reviewed: Yes
ASJC Scopus subject areas: Analytical Chemistry, Biochemistry, Organic Chemistry
Sustainable Development Goals: SDG 3 - Good Health and Well-being
Electronic version(s): http://hdl.handle.net/10033/622357 (Access: Open)
https://doi.org/10.1016/j.chroma.2020.461367 (Access: Closed)