Dextran sulfate protected bFGF from heat and acid inactivation and from proteolytic degradation. The protective effect was stronger than that of hcparin which is known as a stabilizer of bFGF. Dextran ~ulfat¢ and bFGF formed a high molecular weight complex via ionic interaction when mixed together in aqueous solution. The complex was dissociated when the ionic strength was increased and the protective effect was completely abolished. Successive digestion of bFO F with Staphylococcus aureu# V8 protease and pepsin followed by affinity chromatography on an immobilized dextran sulfate column and reversed-phase high performance liquid chromatography yielded three positively charged fragment peptides, Tyr aPhe ~°. Tyrm~-TrpU~ and Tyrt-'a-Leu ~. These results suggest that dextran sulfate stabilizes bFGF by binding close to the putative heparin binding sites of the bFGF molecule. £. coil.derived recombinant bFGF and its genetically engineered acid-stable mutein CS23 (rbFGF-CS23) [6] have been demonstrated to significantly accelerate the healing of chronic duodenal ulcers produced by cysteamine in rats [7]. Oral administration of rbFGF-CS23 caused a significant increase in angiogenesis in the ulcer bed compared with that in untreated control rats [8]. These findings demonstrate the important role(s) of bFOF in the healing of duodenal ulcers and imply a novel pharmacological modulation of gastrointestinal ulcers.Since orally administered rbFGF-CS23 was found to be degraded by the digestive enzymes in the gastrointestinal tract, we examined the use of heparin-related reagents derived from natural and synthetic sources to protect rbFGF-CS23 from proteolytie degradation. Of them, we found that dextran sulfate was most pot~at. We report here that dextran sulfate has high affinity for

Anti-recombinant human basic fibroblast growth factor (rbFGF) antibodies effaeiently inhibited the basal proliferation of bovine pulmonary artery endothelial (BAE) cells. The cell-free extract of BAE cells stimulated the proliferation of bovine capillary endothelial cells and this activity was completely abolished by the antibodies. Furthermore, on heparin HPLC the activity was eluted at exactly the same retention time as that for authentic pituitary bFGF. These observations directly indicate that the BAE cells produce bFGF that stimulates their own basal growth by binding to specific receptors expressed on the cell surface.

No abstract

Two lower-molecular-weight derivatives of recombinant human interferon-gamma (rIFN-gamma) were purified concurrently from a lysozyme-EDTA extract of Escherichia coli cells by immunoaffinity chromatography using a monoclonal antibody (MAb) against a synthetic carboxy-terminal peptide (Lys-131-Gln-146). The two derivatives, 15K rIFN-gamma and 17K rIFN-gamma, were regarded to have been generated at the extraction step. They were successfully separated from each other by using another MAb against the same synthetic peptide with higher binding affinity than the first. The results of protein-chemical analyses indicate that 15K rIFN-gamma and 17K rIFN-gamma lack 15 (Arg 132-Gln-146) and 4 (Arg-143-Gln-146) carboxy-terminal amino acid residues, respectively. All the data suggest that the two derivatives form a noncovalent dimer and that 15K rIFN-gamma binds indirectly to the MAb column via 17K rIFN-gamma.

No abstract