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Purification of the human fibroblast growth factor 2 using novel animal-component free materials
Bolten, Svenja Nicolin Knoll, Anne-Sophie Li, Zhaopeng Gellermann, Pia Pepelanova, Iliyana Rinas, Ursula Scheper, Thomas
Bolten, Svenja Nicolin
Knoll, Anne-Sophie
Li, Zhaopeng
Gellermann, Pia
Pepelanova, Iliyana
Rinas, Ursula
Scheper, Thomas
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2020-08
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Abstract
This paper analyzes the use of animal-component free chromatographic materials for the efficient purifi- cation of the human fibroblast growth factor 2 (hFGF-2). hFGF-2 is produced in Escherichia coli and pu- rified 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 chromatog- raphy were tested as a substitute for heparin. The MMCs were cation exchangers characterized with fur- ther 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 col- umn HiTrap TM Capto TM MMC from GE Healthcare Life Sciences - can be regarded as effective animal- component free alternatives to the heparin - based adsorber.
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en
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0021-9673
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Graded Implants FOR2180
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Attribution-NonCommercial-ShareAlike 4.0 International