Author: Amrit Kannan Major: Biochemistry 

During the course of the 2018-2019 school year, I performed research in the chemistry department under the direction of Dr. TKS Kumar.  Dr. Kumar focuses on a set of naturally-occurring proteins known as Human Fibroblast Growth Factors, or FGFs.  My specific project is to develop a heterodimer version of the two most common variants of FGF, FGF1 and FGF2, to form one protein known as FGF1-FGF2.  The underlying reasoning for making such a protein is due to the fact that FGF1 and FGF2 are quite different from each other; for example, while one is integral in angiogenesis, the other is critical in wound healing.  Making one protein that is multifaceted in its abilities would increase the effectiveness of it, and help the lives of people needing cures to a variety of ailments around the world. 

My project can be defined as one that has been very promising, but wrought with a combination of challenges.  My mentors and I knew that making such a large, unusual protein would come with its set of challenges, and through the course of my time working on this project, these challenges have definitely presented themselves.  Two major problems have slowed the progression of my project: low yields and degradation.  These problems are related to one another, and measures have been taken to remedy them.  Upon receiving low yields, my mentors and I began to use a protease inhibitor cocktail, which negated the effect of enzymes that catalyze amino acids, inherently increasing the viability of our protein.  Next, we switched to using a different host bacterium, which packages its proteins more carefully than the one we had been using.  Finally, we switched to using a different growth media, which further enhanced bacterial and protein growth.  Through these meticulous changes in our methodology, our protein yield increased to more acceptable quantities.  However, we still have a way to go.  In addition to these experiments, further work has been done this semester to purify and characterize the less-stable wild type form of FGF used in the heterodimer, FGF2.  Several purifications and characterization experiments have been performed on this mutant in order to glean more information about it, leading to a potentially more stable version of FGF1-FGF2. 

Through these challenges encountered in my lab, I have learned a lot about myself and how I deal with success, as well as setbacks.  Knowing this project has big implications and believing in my abilities to conquer the challenges I encounter, I have been able to persevere and remain enthusiastic and optimistic with the help of my mentors.  I know that my perseverance will pay off in the future with a potential patent and publication from this project. 

For this summer and upcoming semester, my primary goal is to continue to express the heterodimer in large quantities by making further changes in our expression and purification protocol.  Upon achieving this, I will continue to test its stability by subjecting it to circular dichroism and fluorescence spectroscopy experiments, as well as digestions using enzymes such as trypsin and thrombin.  I will finally test its bioactivity (ability to assist in cell proliferation) and will subject it to NMR spectroscopy to further determine its exact molecular structure.