Date of Graduation
Master of Science in Cell & Molecular Biology (MS)
Robert Beitle, Jr.
Second Committee Member
Collagen Binding Domains, E. coli hosts, fed-Batch, Fibroblast Growth Factor, Fusion Protein
Delivering effective and non-toxic doses of bioactive materials that can aid in activating tissue regeneration to wounded tissue has proven to be an enormous challenge. This study was designed to produce a potential therapeutic recombinant protein by fusing two collagen binding domains to basic fibroblast growth factors (bFGF) through a collagenase cleavage site linker, so it can release the bFGF in a wound site by the action of this enzyme. The novel fusion protein was expressed in Escherichia coli BL-21 (E. coli) using traditional flask shaker and fed-batch cultivation. Cell lysate was purified by FPLC using Immobilized metal affinity chromatography and heparin heparin-affinity chromatography. Results showed that the desired fusion protein was expressed, but different truncated products were also evident including a protease within E.coil may indeed recognize the designed collagenase cleavage site sequence.
Aljewari, H. (2019). Production and Purification of Basic Fibroblast Growth Factor Fused to two Collagen Binding Domains expressed in E. coli BL21 using Flask and fed-Batch. Theses and Dissertations Retrieved from https://scholarworks.uark.edu/etd/3521