Date of Graduation


Document Type


Degree Name

Master of Science in Cell & Molecular Biology (MS)

Degree Level



Biological Sciences


Robert Beitle, Jr.

Committee Member

Josh Sakon

Second Committee Member

Christa Hestekin


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.