Jamie Schnarrs
Pronouns: They/Them
Research Mentor(s): Matthias Truttmann
Research Mentor School/College/Department: molecular and integrative physiology / Medicine
Program:
Authors: Jamie Schnarrs, Asa Huffaker, Sarah Tabler, Mary E. Skinner, Matthias C. Truttmann, Matthias Truttmann
Session: Session 4: 1:40 pm – 2:30 pm
Poster: 53
Abstract
Nanobodies are antibody fragments derived from heavy-chain-only antibodies that are uniquely produced by camelids and sharks. They are a growing area of scientific interest as promising alternatives to conventional antibodies. Antibodies are expensive to produce, require cold storage, and are poor candidates for treatments of neurological pathologies because they are unable to penetrate the blood-brain barrier. Nanobodies are much smaller than antibodies, are temperature stable and possess the potential to penetrate the blood-brain barrier. Nanobodies are far more cost-effective to produce. Many unique nanobody sequences have already been identified, but there are an abundance of promising therapeutic targets for nanobodies that have not yet been established. We aim to not only discover more sequences, but also learn how to optimize their production. Several proteins were selected as promising nanobody target candidates and injected into a llama biweekly, totaling six injections, to induce nanobody production. Lymphocytes were isolated from the llama’s blood and total RNA was then extracted. From there, cDNA was generated and used for amplification of the nanobody binding domains. The amplification products were transformed into a phagemid vector and a phage panning assay was conducted. This allowed us to screen individual E. coli colonies for potential nanobody sequences targeting our candidate proteins. We have identified several unique nanobody sequences that target a variety of proteins, many of which were successfully expressed and purified. Sequences that were not successful in production undergo individual troubleshooting through procedural alteration and experimentation. In the future, we will continue to screen for more unique sequences, as well as alter and optimize our procedures to better future research.
