Considerations on the flow configuration of membrane chromatography devices for the purification of human basic fibroblast growth factor from crude lysates

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.

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

“…The biological activities of heparin are associated with its interaction with various proteins, which have led to its use in protein purification in biotechnological applications [6,7] . Diverse biologically active heparin-binding proteins, such as the human fibroblast growth factor 1 (hFGF-1) [8] , hFGF-2 [9,10] , the human bone morphogenic protein 2 (hBMP-2) [11] , the serine protease throm-bin [12] , and the glycoprotein anti-thrombin III (ATIII) [13] , can be successfully purified by heparin affinity chromatography in a very effective and simple process [7,[14][15][16] . However, heparin is an animal-derived material, which is most frequently obtained from bovine or porcine tissue.…”

Similarly sized protein separation of charge‐selective ethylene‐vinyl alcohol copolymer membrane by grafting dimethylaminoethyl methacrylate

“…Membrane chromatography has been widely used for protein purification separation due to its good penetration, fast mass transfer, lower pressure drop, and easier to scale up, that is, gelatin pre‐concentration and enzymes extraction . One of the important filtration principles of membrane is the size exclusion, but the applications of traditional membrane for separating similarly sized proteins are still limited because the pore size of membrane is constant once prepared and the sieving/selectivity property of membrane is uncontrollable .…”