After declaring chemical engineering as my major, I began looking for research opportunities that interested me. I looked through a brochure that discussed the research of each chemical engineering professor, and Dr. Bundy’s cell-free synthetic biology research with applications in biocatalysis and cancer research fascinated me. I had previously taken a class in molecular biology, and the work the Bundy Biotech Lab was doing was right up my alley.
I met with Dr. Bundy and signed up to start doing research for credit. Conner Earl mentored me and helped me get a feel for what the lab was doing. We soon started working on a project attempting to express a bispecific T-cell engager (BiTE) in a cell-free environment. BiTE consists of two anti-body fragments bonded together. These sing-chain variable fragments (scFvs) are each specific to a different target. One scFv targets T cells, and the other targets cancer cells. In the early stages of cancer, T cells can attack cancer cells. However, as cancer progresses, the cancer cells mutate and gain the ability to elude T cells. BiTE brings the T cell to the cancer cell so it can attack it by binging first to the T cell with one scFv region and then to the cancer cell with the other scFv region.
David Wood’s lab had developed the specific BiTE that we were using previously and had sent us a vector containing the gene encoding for their protein. We were trying to express it in a cell-free environment to take advantage of the benefits of not having to lyse open cell walls to extract the protein and of avoiding the potential undesired variables associated with the cytotoxic environment inside of a cell. These benefits could lead to potential high-throughput production of BiTE for use in fighting cancer.
Along the way Conner, the other undergraduates, and I were able to hone our learn valuable laboratory techniques and improve our ability to design experiments. We learned how to pipette with precision, use a microcentrifuge and an autoclave, perform PCR, make and run gels using gel electrophoresis, take a NanoDrop OD reading, prepare amplified DNA for BigDye Sequencing, do a maxiprep and a miniprep, prepare cell extract, and make glycerol stocks. With this knowledge, we could troubleshoot problems that arose with expressing BiTE. Unfortunately, despite all our efforts,
Writing a Review on CFPS Biosensors
After I had worked in the Bundy Biotech Lab for one year, Dr. Bundy announced that Mehran, a graduate student from Iran, would be in charge of writing a review article about cell-free synthetic biology due in the next six weeks. Knowing that Mehran was only a second-semester graduate student and would likely have difficulty writing a review article in English, I volunteered to help Mehran with the article.
Writing a review article, which includes over 60 citations and requires novel insight on a minimally explored topic, takes an enormous amount of time. As a result, I asked Dr. Bundy if we could meet with him immediately following the meeting. Dr. Bundy told us that we would specifically be reviewing cell-free biosensors and gave us several articles to look at, an outline for the introduction, and a general outline for the paper.
Over the next month, Mehran and I worked closely together to gather resources, discuss research, and outline the paper. We oversaw two other undergraduates, Andrew Nelson and Hayley Ford, as they helped research articles. Andrew wrote the introduction based on an outline that I made, and Hayley designed beautiful figures based on some sketches that I made.
When it came time to write the main body of the paper, Mehran suggested that he write one half and I write the other. Periodically, we looked over each other’s writing and offered suggestions for improvement. When the manuscript was ready, we proofread what we had written and made sure that the citations were formatted correctly.
This experience helped me learn how to clearly write about complex ideas in the form of a rhetorical argument, collaborate with another author to develop a manuscript, and prepare a manuscript for submission to an academic journal. Technical writing requires a consistent, concerted effort and teamwork. This project made me a better leader and gave me greater confidence that I can accomplish difficult things.