Multiplexed Antigen Attachment Using DNA Nanoparticles for Platform Vaccine Development

Due to the sensitive nature of my research this summer, I am unable to provide my full abstract at this time. I am happy to speak further about this or direct you to Dr. Veneziano with any further questions.

Vaccines have been used for over 200 years, starting when Edward Jenner first administered the smallpox vaccine in 1798. Despite the incredible positive impact that vaccines have had on public health for the past 2 centuries, if you had asked me or my colleagues a year ago, none of us would have ever guessed that vaccines would be the global hot topic they are now. There are many different ways to create a vaccine, but the novel approach that I have been working on this summer involves using brand new DNA nanoparticle technology as a biocompatible and versatile vector to safely deliver antigen and produce an immune response so the pathogen can be recognized by the body  in the future. The process of creating the DNA nanoparticles that are used as a base for our vaccine technology begins with producing the scaffold for the desired 3 dimensional structure using asymmetric polymerase chain reaction (often referred to as aPCR).  After adding all of the necessary primers, buffers, template, and DNA, I run the mixture through aPCR and it comes out as a mix of both double stranded DNA and the desired single stranded DNA.  I then isolate the ssDNA by running it through gel electrophoresis in an agarose gel that I have prepared ahead of time. This process of gel electrophoresis separates out the ssDNA from the double stranded DNA.  The one band shown at the start of the experiment turns into two separate strands, the top band being the dsDNA, and the bottom is the lighter ssDNA that travels further through the gel.  I then manually cut out the ssDNA using a razor blade and begin the purification process.  The purification process involves heating the gel in a liquid until it is completely liquified, and then running it through a centrifuge multiple times with a washing buffer and eventually an elution buffer.  All of the ssDNA gets bound on a silica matrix that is made specifically for this process, and the elution buffer rinses the purified ssDNA that results from this process into a separate test tube so that I can use it in the folding process. At this point, our scaffold has been created and it is combined with the correct staple strand and put through a 12 hour folding process known as “annealing” which involves very specific slow decreases and drops in temperature to aid in the folding process.  The last step is to attach the antigens we will use to the DNA nanoparticles using specific chemical attachment techniques, and then I run the particles through surface plasmon resonance which allows me to measure how well the nanoparticles bind based on antigen positioning and type. This technique is superior to other methods of antigen attachment because while current vaccine particles allow for attachment, they cannot control the precise location or stoichiometry of the attachment. What I have been doing in the lab allows for complete control over the stoichiometry and placement of the proteins, in addition to being biocompatible and biodegradable, and it has been proven to work under similar conditions (using the HIV virus). Prior to my research, this method has been tested once using one specific antigen on the surface of a DNA nanoparticle, but what the groundbreaking technology of DNA origami has to offer is still just beginning to be explored. It is my hope that the results of my research this summer will not only satisfy my own personal curiosity, but that it will open countless doors for vaccine development and the practical, worldwide use of DNA origami in the future.