UNC-33 and its human homolog, CRMP2 (Collapsin Response Mediator Protein-2), have been demonstrated to be involved in neurodevelopment as well as neurodegenerative disorders, primarily Alzheimer’s Disease. However, the physiology and interactions of these associations are vague. In order to further understand UNC-33/CRMP2, our group decided to use molecular biology and work toward the production of polyclonal antibodies specific to C. elegans UNC-33. To do this, we utilized the GST tag Gene Fusion System and produced two antigens- UNC-33 amino acid 48 to 212 and UNC-33 amino acid 48 to131 (UNC-3348-212 and UNC-3348-131). During this process, parameters were developed for the efficient expression and purification of these polypeptides. Once an effective protocol was established, GST fused UNC-3348-212 and UNC-3348-131 were expressed, purified, and tested for purity multiple times. Overall, these procedures resulted in the production of 3.72 mg and 2.10 mg of GST fused to UNC-3348-212 and GST fused to UNC-3348-131, respectively. Currently, these purified polypeptides are being injected into laboratory animals for the generation of polyclonal antibodies for UNC-33 research.
Abbott, Matt; Fuller, Jacob; Howe, Mason; Caniglia, Michael; and Holgado, Andrea
"Generating Caenorhabditis elegans UNC-33 antigens to be used for the Synthesis of Polyclonal Antibodies,"
SWOSU Journal of Undergraduate Research:
Vol. 1, Article 3.
Available at: http://dc.swosu.edu/jur/vol1/iss1/3