Details
| Original language | English |
|---|---|
| Article number | 461367 |
| Journal | Journal of Chromatography A |
| Volume | 1626 |
| Early online date | 23 Jun 2020 |
| Publication status | Published - 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
- Chemistry(all)
- Analytical Chemistry
- Biochemistry, Genetics and Molecular Biology(all)
- Biochemistry
- Chemistry(all)
- Organic Chemistry
Sustainable Development Goals
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In: Journal of Chromatography A, Vol. 1626, 461367, 30.08.2020.
Research output: Contribution to journal › Article › Research › peer review
}
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 -