Abstract

Heparin Enhancementof VEGF Binding did not displace '"1-VEGF in competition studies?Recombinant human bFGF was produced in bacteria using the T7based expression vector pet-8C (21) (kindly provided by Dr. F. W. Studier, Brookhaven National Laboratory, Upton, New York) into which the full length human bFGF cDNA was ligated between the NcoI and the BamHI sites.Bacteria (BL21(DE3)) containing the pLys-S plasmid (21) were lysed by the method of Studier (21), and the bFGF was purified from the lysate by heparin-Sepharose affinity chromatography (22).Heparin-Sepharose was purchased from Pharmacia (Uppsala, Sweden).'2sI-Sodium was purchased from Amersham (Buckinghamshire, England).Bacterial heparinase was obtained from Ibex Technologies (Montreal, Canada).Bovine lung (H-9133) and porcine intestinal mucosa-derived heparin (H-7005) were purchased from Sigma.The porcine mucosa-derived heparin was generally used, unless otherwise stated.Heparin fractions with high (HA) and low (LA) affinity to antithrombin I11 were separated using an antithrombin-Sepharose column as described (23).This heparin was purchased from Hepar