Modifying the Protein Construct for Nanoparticle Adhesion

Undergraduate #2
Discipline: Biological Sciences
Subcategory: Biomedical Engineering

Jessica Leal-Cruz - University of California, Irvine

Site-specific targeting has become an essential need in the field of cancer treatment and detection. Nanoparticle carriers connected to DNA proteins for adhesion have the potential to become the necessary tool to achieve this goal. Our current construct, however, has shown very little differentiation in selectivity between diseased cells and healthy cells. By creating variations of the construct, we can observe and compare their binding kinetics and selectivity. In this case the Soratse tag and PYF vector were adapted to the construct. Plasmids: pDrive- Sortase and PYF-4420 were amplified and digested in order to ligate the Sortase insert and PYF vector. Positive cell growths on plate and positive diagnostic agarose gels provided evidence that the PYF+Sortase Tag DNA plasmid were ligated successfully. This gives further motivation to use the Sortase tag with other vectors for comparison of binding kinetics. Manipulating protein
constructs could give a better understanding of how to improve selectivity for targeted antibodies.

References: American Cancer Society. Cancer Treatment and Survivorship Facts & Figures 2014-2015. Atlanta: American Cancer Society (2014). Haun, J. B. and Hammer, D. A. Quantifying Nanoparticle Adhesion Mediated by Specific Molecular Interactions. Langmuir 24 (2008). Haun, J. B. et al. Using Engineered Single-Chain Antibodies to Correlate Molecular Binding Properties and Nanoparticle Adhesion Dynamics. Langmuir 27, 13701-13712 (2011). Warden-Rothman, R. et al. Sortase-Tag Expressed Protein Ligation: Combining Protein Purification and Site-Specific Bioconjugation into a Single Step. Analytical Chemistry 85, 11090-11097.

Funder Acknowledgement(s): Thank you to Kika Friend, CAMP Director for the opportunity to do research at UCI and to Professor Jered Haun for his support and guidance through this research. This work is supported by the Louis Stokes Alliance for Minority Participation (LSAMP)/ California Alliance for Minority Participation (CAMP) at the University of California, Irvine.

Faculty Advisor: Jered Haun,