WIP

Cultured various human cancer cell lines for in vitro testing of ADCs to assess their cytotoxicity and the strength and specificity by which they bind targets. Read out my data using luminescence reagents, plate readers, and flow cytometry. Also investigated ADC-instigated protein degradation with western blotting. Carried out all my own data analysis and compiled final results and procedures together in Benchling entries.Also helped with lab chores such as glassware sterilization. Read literature for active participation in literature discussion meetings.

Assisted colleagues with their experiments by culturing B16 melanoma cells for them, necropsying mice and isolating lymphocytes from their tissues, assisting with mouse surgery, and carrying out injections of cells and drugs into mice. Also often stained and carried out flow cytometry on the lymphocytes isolated from mice. For one project I isolated RNA from mouse tissue and analyzed it with RT-qPCR to generate gene expression data. In another project I carried out IHC on mouse lymph nodes from embedding to sectioning to staining and imaging. On a more routine basis, I propagated and harvested hybridoma culture to replenish lab monoclonal antibody stocks. I also ran the Turk lab mouse colony. Occasionally replenished B16 stocks by serial in vivo passage and subsequent in vitro expansion.For another project I isolated PBMCs from human blood using Ficoll-Paque and then further isolated naive B cells. I used these naive B cells to test various combinations of interleukins for their ability to induce isotype switching, and read the results out with qPCR and flow cytometry.Also carried out ordering, expense submission, drug tracking, and other administrative tasks.

I received orders for viruses and planned my production schedule accordingly. I applied my cell culture techniques to produce adenovirus and AAV in the correct volume, concentration, container, and packaging. I also performed some of the required QC testing on my products. Regularly carried out density ultracentrifugation. During my time at ABM I also helped update and otherwise improve company SOPs and assisted in some internal R&D.

As a Forensic Technologist Understudy I learned how to examine exhibits visually while paying extreme attention to detail and how to use chemical and biological tests in accordance with protocols. I also recovered and preserved biological material such as hair, blood, and semen, as well as non-biological materials such as glass and fibers in accordance with protocols.I recorded my observations and exhibit chain of custody in LIMS software and processed these notes into reports using LIMS-related software. I performed quality control testing on critical reagents to maintain rigorous QA standards. I also performed in-depth file review and assisted with administrative tasks.

I propagated a virus in mammalian cell culture and extracted it in bulk with sonication and freeze-thaw technique. I further purified some of this crude lysate swine virus stock with liquid-liquid extraction and/or a sucrose gradient and then characterized the resultant virus preparation using immunofluorescence, qPCR, and electron microscopy. The purified virus was used in ELISAs to screen animal sera for highly reactive antibodies.I also coupled reactive latex beads to that same virus and used the resulting virus-coated beads to extract antibodies from swine sera. These extracted swine antibodies were then used in phage display in an attempt to find peptides that mimic the antigenicity of the swine virus. This project gave me a great deal of expertise in phage display techniques such as blue-white screening.

I interacted with the public for the purpose of convincing individuals to either fill out a survey or to allow us to email the survey to them. This position required initiative and good interpersonal skills. I had no problem with approaching strangers. I only stopped working in this position because I received a job offer from my old laboratory in March, and I wanted to get back to science. I continued working for PureGold on weekends until the end of March.

I used reverse genetics to propagate and purify a specific strain of a swine virus. Specifically, I transfected swine cells in culture with a plasmid construct containing the viral genome. These transfected cells were passaged multiple times to replicate more virus. I extracted the virus from the cell culture using sonication and the freeze-thaw technique. The TCID50 of the virus stock was evaluated using titration and its purity from contamination with a closely related virus was evaluated using qPCR. I also used immunofluorescence to analyze the in vitro infection kinetics of the virus. I also had the pleasure of training a new lab member during this time.

Please see Education.

I expressed and purified recombinant viral proteins for use in ELISAs to screen animal sera for antibodies to these viral proteins. I expressed these proteins in bacteria and extracted them in denaturing conditions to maximize yield, and then purified them using IMAC. Mass-spec and immunoblotting were used to confirm the presence of the protein of interest and its purity was evaluated using Coomassie and silver stain. The fractions containing semi-purified viral protein were pooled and used as antigen in the ELISAs, which were first optimized along the parameters of antigen amount, sera dilution, and secondary antibody dilution.

Lab Assistant: prepared reagents and media as required. Cleaned and sterilized glassware and sterilized other consumables. Calibrated pH meter daily. Loaded samples onto SDS-PAGE gels, ran and stained them, and then dried these gels. Stocked supplies and chemical reagents as required.Honour Student: please see Education.

Duties included standard QA/QC testing of equipment and media/reagent preparation. Logged lab temperatures daily. I extracted PSP toxin from shellfish and logged PSP test results into a central database. Calibrated thermometers and conducted productivity testing of media stocks. Assisted in SOP audit.