ABSTRACT
This chapter describes the use of heparin affinity high-performance liquid chromatography (HPLC) in the purification of fibroblast growth factors from tissues and cells. A wide range of proteins bind strongly to heparin, a highly anionic glycosaminoglycan, and affinity chromatography on heparin covalently linked to a matrix has often been used as a purification step for biologically active heparin-binding proteins. Two of these heparin-binding endothelial cell growth factors turned out to be acidic fibroblast growth factor (aFGF) and basic fibroblast growth factor (bFGF), polypeptides originally isolated from brain. Heparin affinity chromatography is a technique which is fairly specific for the purification of aFGF and bFGF. Conventionally, heparin-Sepharose affinity chromatography has been used for the purification of aFGF and bFGF. Heparin affinity HPLC has several advantages. Since the HPLC column matrix generally is more regularly packed and the gradient is more accurate, resolution is higher and results are highly reproducible.
