Details
| Original language | English |
|---|---|
| Pages (from-to) | 697-705 |
| Number of pages | 9 |
| Journal | Engineering in life sciences |
| Volume | 16 |
| Issue number | 8 |
| Publication status | Published - 1 Jul 2016 |
Abstract
Membrane chromatography possesses numerous advantages such as operation at high flow rates, low back pressure, ease of handling and scale up, which make the membrane adsorber process a viable alternative to conventional packed column chromatography. A purification process for the isolation of human recombinant basic fibroblast growth factor (FGF-2) based on membrane chromatography was investigated using devices with different flow configurations. In the first process, the FGF-2 capture step was performed with an axial flow device, while the alternative method achieved direct capture of FGF-2 from unclarified cell lysate with a tangential flow device. In both processes, FGF-2 purities exceeded 82% and the purified cytokine displayed high biological activity. Binding capacity (BC) from fermentation broth of the axial flow device was 28 mg/mL. This was 50% higher than the BC obtained with the tangential flow device under particle-free supernatant conditions (18 mg/mL) and 150% higher compared to the BC achieved with unclarified cell lysate (11 mg/mL). While membrane chromatography in tangential flow mode omits clarification and thus reduces the number of stages in the downstream process, it displays lower peak resolution and leads to a lower overall process yield.
Keywords
- Axial flow device, Direct capture, FGF-2, Membrane chromatography, Tangential flow device
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Biotechnology
- Environmental Science(all)
- Environmental Engineering
- Chemical Engineering(all)
- Bioengineering
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In: Engineering in life sciences, Vol. 16, No. 8, 01.07.2016, p. 697-705.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Considerations on the flow configuration of membrane chromatography devices for the purification of human basic fibroblast growth factor from crude lysates
AU - Gieseler, Gesa
AU - Pepelanova, Iliyana
AU - Meyer, Annette
AU - Villain, Louis
AU - Beutel, Sascha
AU - Rinas, Ursula
AU - Scheper, Thomas
PY - 2016/7/1
Y1 - 2016/7/1
N2 - Membrane chromatography possesses numerous advantages such as operation at high flow rates, low back pressure, ease of handling and scale up, which make the membrane adsorber process a viable alternative to conventional packed column chromatography. A purification process for the isolation of human recombinant basic fibroblast growth factor (FGF-2) based on membrane chromatography was investigated using devices with different flow configurations. In the first process, the FGF-2 capture step was performed with an axial flow device, while the alternative method achieved direct capture of FGF-2 from unclarified cell lysate with a tangential flow device. In both processes, FGF-2 purities exceeded 82% and the purified cytokine displayed high biological activity. Binding capacity (BC) from fermentation broth of the axial flow device was 28 mg/mL. This was 50% higher than the BC obtained with the tangential flow device under particle-free supernatant conditions (18 mg/mL) and 150% higher compared to the BC achieved with unclarified cell lysate (11 mg/mL). While membrane chromatography in tangential flow mode omits clarification and thus reduces the number of stages in the downstream process, it displays lower peak resolution and leads to a lower overall process yield.
AB - Membrane chromatography possesses numerous advantages such as operation at high flow rates, low back pressure, ease of handling and scale up, which make the membrane adsorber process a viable alternative to conventional packed column chromatography. A purification process for the isolation of human recombinant basic fibroblast growth factor (FGF-2) based on membrane chromatography was investigated using devices with different flow configurations. In the first process, the FGF-2 capture step was performed with an axial flow device, while the alternative method achieved direct capture of FGF-2 from unclarified cell lysate with a tangential flow device. In both processes, FGF-2 purities exceeded 82% and the purified cytokine displayed high biological activity. Binding capacity (BC) from fermentation broth of the axial flow device was 28 mg/mL. This was 50% higher than the BC obtained with the tangential flow device under particle-free supernatant conditions (18 mg/mL) and 150% higher compared to the BC achieved with unclarified cell lysate (11 mg/mL). While membrane chromatography in tangential flow mode omits clarification and thus reduces the number of stages in the downstream process, it displays lower peak resolution and leads to a lower overall process yield.
KW - Axial flow device
KW - Direct capture
KW - FGF-2
KW - Membrane chromatography
KW - Tangential flow device
UR - http://www.scopus.com/inward/record.url?scp=84981306992&partnerID=8YFLogxK
U2 - 10.1002/elsc.201600006
DO - 10.1002/elsc.201600006
M3 - Article
AN - SCOPUS:84981306992
VL - 16
SP - 697
EP - 705
JO - Engineering in life sciences
JF - Engineering in life sciences
SN - 1618-0240
IS - 8
ER -