Discipline: Chemistry and Chemical Sciences
Regine Hilary Leopoldine Vincent - Indiana University Bloomington
Co-Author(s): Austin Reilly, Indiana University, Bloomington, IN, 47405, USA Jasper Dittmar, Indiana University, Bloomington, IN, 47405, USA Barry D. Stein, Indiana University, Bloomington, IN, 47405, USA Lyudmila M. Bronstein, Indiana University, Bloomington, IN, 47405, USA
We focus on the development of thermally stable iron oxide nanoparticle-based vaccines for the Human Papilloma Virus (HPV-16) through the assembly of L1 protein pentamers (capsomers) or L2 epitope oligopeptides on the surface of functionalized iron oxide nanoparticles (NPs). Although several effective vaccines exist against HPV-16, such vaccines are often expensive and lack thermal stability, thus presenting delivery and distribution problems for the developing nations that utilize these vaccines. NP functionalization was conducted by utilizing three types of PEGylated phospholipids with thiol, carboxyl, and amino terminal groups. Protein attachment with the SH and NH2 functionalized NPs was achieved through the use of an NHS crosslinker and covalent attachment. The COOH functionalized NPs utilized an electrostatic self-assembly process to conduct HPV protein attachment to the nanoparticle surface. Attachment conditions were studied by varying pH and ionic strength of the solution, as well as by modifying protein to nanoparticle to crosslinker ratios.
Upon functionalization, transmission electron microscopy (TEM) was employed to observe the efficiency of phospholipid coating. After protein attachment, immunogold staining as well as TEM were utilized to determine the presence of virus protein on the nanoparticle surface. We demonstrate that COOH-functionalized NPs are the most efficient, realizing HPV-NP constructs. For amino- and thiol terminated NPs, the assembly conditions should be further modified to achieve more efficient attachment. Additionally, results show that efficiency of NP-Oligopeptide attachment is inconclusive. Continued work will involve optimizing protein attachment by further modifying attachment conditions and by adapting conditions to promote NP-Oligopeptide construct formation.
Funder Acknowledgement(s): This work was made possible by the Indiana University Department of Chemistry, the Hutton Honors College Summer Research Grant, the Raymond Siedle Scholarship, and the Louis Stokes Alliance for Minority Participation. Special thanks to the Indiana University Bloomington Physical Biochemistry Instrumentation Facility for access to instrumentation, and to Aaron Ermel and Brahim Qadadri of the Indiana University School of Medicine for providing HPV capsomers.
Faculty Advisor: Lyudmila Brosntein, email@example.com
Role: I am the primary researcher in this project. My role in the researcher encompassed all that was necessary to carry out the research. My responsibilities included functionalizing and purifying nanoparticles, attaching HPV protein to the functionalized nanoparticles, as well as performing immunogold staining procedures to image the HPV-nanoparticle constructs.