Research Lab Technician
Stony Brook University, Dept. of Structural Biology and Biochemistry
July 2020 – Present
Under the supervision of Dr. Steven Glynn, my primary objective is to develop an optimized expression and purification strategy for a membrane protein resident to the mitochondria called Tafazzin. Tafazzin is a transacylase responsible for remodeling the phospholipid cardiolipin (CL) into its mature form. Mature CL inserts into the mitochondrial inner membrane, stabilizing its curvatures (cristae) and the oxidative phosphorylation complexes which reside there. Several known mutations in Tafazzin result in the enzyme being made inactive or mislocalized, resulting in a disorder known as Barth’s syndrome.
Due to the intrinsic instability of membrane proteins in aqueous solution, there are a variety of bottlenecks to purifying this category of protein and, therefore, only a small percentage of membrane protein structures have been resolved. Our hope is to use our optimized protocol, once complete, to determine the structure of Tafazzin via X-ray crystallography. Having a 3D structure of this enzyme will lay the foundation for a much better understanding of how it functions, why it’s mutant variants don’t, and how we may intervene with medicine to alleviate it.
In addition to this project, I’m tasked with general lab upkeep, stocking solutions, and training new students on laboratory techniques and equipment.
Skills:
- Bacterial cell culture
- Cloning
- DNA extraction
- PCR
- SDS-PAGE
- Western Blot
- Protein purification
- IMAC
- SEC
- IEX
- ÄKTA system
- Image analysis with ImageJ
Manufacturing Technician
Kedrion Biopharma
March 2020 – July 2020
Kedrion Biopharma derives therapeutics from human blood plasma for the treatment of rare disorders such as hemophilia, Hemolytic disease of the newborn (HDN), and immunodeficiencies.
My job as a Manufacturing Technician entailed the initial processing of incoming shipments of plasma from blood donations throughout the region. It was a very team-oriented position that required equal effort from all technicians. After unloading shipments into the facility, we inspected all bottles for deviations and prepared them for transfer to the first sterile processing room. In this next room, we would systematically remove the plasma from their bottles, load them into buckets, and transfer the buckets to a lift that would raise and pour the plasma into thaw tanks.
Next, the plasma would be transferred to various centrifuges and filters where it would be fractionated into its various components for further downstream processing. My team and I were responsible for breaking these equipment down for cleaning/sterilization and resetting them for the next use.
As this was my first experience in the biopharmaceutical industry, I learned a great deal about strictly abiding by Standard Operating Procedures (SOP’s) and current good manufacturing practices (cGMP’s) in an FDA-regulated facility. We had daily, weekly, and monthly cleaning procedures for the entire facility. It was hard work, but very humbling knowing our efforts were helping the company develop orphan drugs for people with life-threating conditions.
Skills:
- Familiarity with following SOPs and cGMP guidelines in an FDA-regulated biopharmaceutical environment
- Meticulous cleaning and sterilization skills
- Teamwork
Volunteer Lab Assistant
Stony Brook University, Dept. of Biomedical Engineering
August 2018 – May 2019
I volunteered at two different labs at SBU while I was attending for my Master’s in biomedical engineering: the labs of Dr. David Rubenstein and Dr. Danny Bluestein.
In Rubenstein’s lab, I learned how to culture mammalian fibroblast cells, electrospin nanofibers for cell scaffolding experiments, and an abundance of information on the physiology and histology of the cardiovascular system.
In the Bluestein lab, I cooperated with two other students to extract platelets from blood and characterize their activation when exposed to a synthetic polymer under physiological sheer stress. I also tested platelet phosphorylation pathways to potentially identify key differences between fetal and adult platelet activities. Both labs extremely insightful for me in terms of practical, hands-on skills and I’m grateful for the people that mentored me during my time there.
Skills:
- Lab safety
- Mammalian cell culture
- Electrospinning
- Microscopy
- Column chromatography
- Centrifuge
- Coulter cell counter
- Flow cytometry