GMP Production of BFGF-Saporin Fusion Protein

Basic fibroblast growth factor (bFGF) receptors are unregulated on the proliferating cells that arepresent in various diseased states, including restenosis, various cancers and various opthamalogicalconditions and syndromes. It has been shown that bFGF coupled to the toxin, saporin inhibits the growthof proliferating cells in both in vitro and in vivo models of these diseases. Using recombinant genetechnology fusion proteins of bFGF-saporin (FGF-SAP) have been expressed in E. coli. At present, asub-optimal protein purification protocol exits that results in milligram quantities of FGF-SAP from gramquantities of starting material. The specific aims of this research are to (1) develop an efficient andexpedient protein purification protocol in order that gram quantities of FGF-SAP can be manufacturedto facilitate preclinical and clinical trials, (2) ensure that this protocol will lend itself to easy scale up andconversion to a GMP process in the following phase of the work and (3) develop a number ofbioanalytical assays to characterize the products which will be used in the validation documentation ofthe GMP process.