TY - JOUR

T1 - Purification of the human fibroblast growth factor 2 using novel animal-component free materials

AU - Bolten, Svenja Nicolin

AU - Knoll, Anne Sophie

AU - Li, Zhaopeng

AU - Gellermann, Pia

AU - Pepelanova, Iliyana

AU - Rinas, Ursula

AU - Scheper, Thomas

N1 - Funding Information: This work was supported by Forschergruppe the research group “ Graded Implants FOR2180 ” and the Cluster of Excellence “Rebirth” EXC62 , both German Research Foundation (DFG).

PY - 2020/8/30

Y1 - 2020/8/30

N2 - 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.

AB - 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.

KW - Animal-component free

KW - Heparin affinity chromatography

KW - Human fibroblast growth factor 2 (hFGF-2)

KW - Mixed-mode chromatography

KW - Pseudo-affinity chromatography

UR - http://www.scopus.com/inward/record.url?scp=85087107558&partnerID=8YFLogxK

U2 - 10.1016/j.chroma.2020.461367

DO - 10.1016/j.chroma.2020.461367

M3 - Article

C2 - 32797846

AN - SCOPUS:85087107558

VL - 1626

JO - Journal of Chromatography A

JF - Journal of Chromatography A

SN - 0021-9673

M1 - 461367

ER -