Inah Bianca Embile

Provide support in developing cell therapies into clinical trials Preclinical safety and biodistribution studies Animal models for chronic conditions and regenerative medicine 2D/3D cell culture and other in vitro assays including protein/DNA/RNA extractions, RT-qPCR and qPCR, ELISAs, Flow cytometry and histology

As part of antibody core, I produce the antibodies needed for research in the department: 1. Animal Handling - administer boosts to mice/rabbits - blood collection - serum titer testing using ELISA - euthanasia/exsanguination - aseptic harvesting of spleen 2. Hybridoma Technology - isolation of PBMCs and immune serum from blood - collection of splenocytes and fusion with myeloma cells - screening of HGPRT positive hybridomas - series of dilutions and cell passaging coupled with ELISAs for selection of specific target antibodies until a final monoclonal antibody product that satisfy particular biochemical/immunological assay requirements (such as for Western Blots or IP) is achieved - expansion from 96w into larger vessels up to bioreactors or roller bottles, ultimately purifying with Protein A/G - Coomassie staining and Western blots to verify protein binding as well as for quality control 3. Apply molecular cloning techniques to determine sequence of hybridomas as alternative - RNA extraction and subsequent formation of ssDNA and then dsDNA using thermocyclers - amplification of regions of DNA using PCR, running DNA through electrophoresis gel, and then isolating and purifying the DNA band to be prepped with specific primers and sent to sequencing for analysis - subject DNA to 5'3' exonuclease activity to prepare it for restriction enzyme editing, which includes the following processes: digestion, ligation and plasmid transformation, and then expanded and isolated for transfection - transient and stable transfections coupled with ELISAs and Western Blots to test viability and success of antibodies and cell lines established Other techniques not mentioned above include: -cell culture techniques: thawing, cell seeding, cell observation, passage of suspension and adherent cells /cell detachment, cell counting, cryopreservation

This position is my first immersion into research, and I couldn’t have joined the Gamez Lab in the Analytical Chemistry Department of TTU at a better time. Then Ph.D candidate, Dr. Gong was embarking on the next chapter of a project she was working on, which was the addition of a laser diode to coupled Low-Temperature Plasma Mass Spectrometry (LTP-MS) and Thin Layer Chromatography (TLC), an already highly sensitive method of detecting biomolecules - so I got to be involved right at the inception of a project. Lectures in school and the pages of sciences books as I grew up came alive as I became more familiar with the scientific method: Asking questions, making observations, drawing hypotheses, designing and executing experiments, and drawing conclusions that can then be used to go back to the drawing board, to redesign and make adjustments to the process. Other practices in research that I got to experience included periodic updates within the group, which allowed for collaborative suggestions on our project as well as theirs; attending a Chemistry convention which allowed for exploration of other studies; and presenting our project at a conference at TTU.