Relative retention of the fibroblast growth factors FGF-1 and FGF-2 on strong cation-exchange sorbents.
The isocratic retention of two heparin-binding fibroblast growth factors, FGF-1 (acidic FGF) and FGF-2 (basic FGF), was compared on a set of six preparative strong cation-exchange adsorbents. The FGFs comprise a solute pair that are structurally equivalent, yet differ in protein parameters of potential importance in cation-exchange chromatography, such as isoelectric point, net charge, and the number and distribution of basic amino acids. The cation-exchange adsorbents comprise a diverse set of materials in common use for protein purification, with physical and chemical properties that have been characterized and described previously. Isocratic k' values for the two proteins obtained on each adsorbent at several different [NaCl] are compared with one another and with corresponding data for hen egg lysozyme, which is also strongly retained on cation-exchangers. Of the six adsorbents examined, three showed strong retention of both FGFs, with equivalent k' values for FGF-1 and FGF-2. Three others, which showed weaker overall retention for the FGF pair, showed much larger retention differences between FGF-1 and FGF-2. The trends in retention order among the stationary phases are very similar to those seen previously with other unrelated proteins. However, retention differences between the two FGFs, and between the FGFs and lysozyme, do not correlate well with simple charge properties such as net charge, indicating, as in some previous studies, the importance of local regions on the protein surface in determining retention. These observations are interpreted in terms of the structural features of the proteins and the physicochemical properties of the adsorbents.
DePhillips
Peter
P
Merck Research Laboratories, Sumneytown Road, West Point, PA 19486, USA.
Lenhoff
Abraham M
AM
D000327:Adsorption; D002411:Cation Exchange Resins; D002852:Chromatography, Ion Exchange / Q000379:methods*; D016220:Fibroblast Growth Factor 1 / Q000737:chemistry*; D016222:Fibroblast Growth Factor 2 / Q000737:chemistry*
D002411:Cation Exchange Resins; D016222:Fibroblast Growth Factor 2; D016220:Fibroblast Growth Factor 1