Progress 01/01/24 to 12/31/24
Outputs
Target Audience:Two presentations were given: one in Nanoscale Science and Engineering for Agriculture and Food Systems Gordon Research Conference (June 23-28, 2024) and the other in CeZAP Infectious Disease Research Symposium (Virginia Tech) (Oct. 11, 2024). The target audience for the Gordon Research Conferences are researchers from accross the country in the use of nanotechnology in agriculture and food. The target audience for the VT CeZAP symposium are immunologists in vaccine development. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?One graduate student, Kristina Ly, gradauted with a MS degree in October 2024. One graduate student, Qiaoqiao Ci, is currently working on the project. Ci was the leading author for two conference presentations, and Ly was a co-author for both.Both students had opportunities to work with undergraduate students to develop their mentoring ability. How have the results been disseminated to communities of interest?Our conference presentations, particularly the on in Gordon Research Conference, is widely accessible for whomever is interested. The MS thesis has been digitally deposited in VT library, which is searchable and accessible forinterested audience. What do you plan to do during the next reporting period to accomplish the goals?We plan to construct the nanovaccines and to condut animal expreriments planned in Objective 2.
Impacts
What was accomplished under these goals?
Objective 1 is almost completed. We ran into difficulty of expressing the full length S protein of PEDV. We have changed to express the S1 domain and the C-terminal Domain (CTD) of the S1 protein as the targeted antigens for vaccine construction. CTD has been successfully produced, and we are in the process of producing S1. Once these two proteins are ready, we will move to objective 2 and 3.
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2024
Citation:
Qiaoqiao Ci, Kristina Ly, Yuanzhi Bian, Riley DeHority, Debra Walter, XJ Meng, Chenming Zhang. Development of a safe and effective nanoparticle-based vaccine against porcine epidemic diarrhea virus. (Poster) 2024 Nanoscale Science and Engineering for Agriculture and Food Systems Gordon Research Conference. June 23-28, 2024. Southern New Hampshire University in Manchester, NH.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2024
Citation:
Ci, Q., Ly, K., Bian, Y., DeHority, R.A., Walter, D.L., Meng, X.J., Zhang, C. Development of a safe and effective nanoparticle-based vaccine against porcine epidemic diarrhea virus. 2024 CeZAP Infectious Disease Research Symposium. Oct. 11, 2024. Blacksburg, VA.
- Type:
Theses/Dissertations
Status:
Published
Year Published:
2024
Citation:
Kristina Ly. Expression and Purification of the C-Terminal Domain of Porcine Epidemic Diarrhea Virus (PEDV) Spike Protein. (MS thesis)
|
Progress 01/01/23 to 12/31/23
Outputs
Target Audience:One manuscript was published, and the main audiences are researchers for animalvaccines or other vaccine scientists. In addition, two presentations were given: one inIMMUNOLOGY2023 (The American Association of Immunologists), one inUSDA National Institute of Food and Agriculture, A1511 Nanotechnology for Agriculture and Food Systems Annual meeting. The target audience for the conferences are immunologists in vaccine developmentandresearchers from accross the country in the use of nanotechnology in agriculture and food, respectively. Changes/Problems:The major change was the target antigen to be used for PEDV nano-vaccine construction. We attempted to express the full length spik protein of PEDV, but the proteinwas proven to be very difficult to be expressed by E. coli. We decided to change to the S1 subunit of the protein, which contains the receptor binding domain and has been shown to be a promising vaccine target. The expression of the S1 subunit is underway. What opportunities for training and professional development has the project provided?Two graduate students worked on this project, one focusing on identifying the adjuvants that could enhance the immunogenicity of the vaccine, and the other focusing on expressing the target antigens for PEDV vaccine construction. One student gave a presentation in a technical conference. Both students had opportunities to work with undergraduate students to develop their mentoring ability. How have the results been disseminated to communities of interest?Yes. The manuscript was published on line. It should be able to reach all interested researchers. Moreover, we gave presentations in two conferences, both attended by researchers in academia and industries (for vaccine development). What do you plan to do during the next reporting period to accomplish the goals?The main focus of the next reporting period is to obtain the target PEDV antigen for vaccine construction. Originally, we focused on the expression of the full length of the spike protein of PEDV. After months of research, the protein was shown to be very difficult to be expressed. Now, we are focusing on the expression of the S1 subunit of the S protein. The S1 subunit contains the domain where the virus interacts with the susceptible cellsand is a promising vaccine antigen. We strive to complete the expression and purification of the protein and test the immunogenicity of the protein.
Impacts
What was accomplished under these goals?
The major goal that was accomplished in thisreporting period is to identify molecular adjuvants that can effectively activate dendritic cells and subsequently the immune system (Objective 1).
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2023
Citation:
Yuanzhi Bian, Debra L. Walter, Chenming Zhang. Efficiency of interferon-? in activating dendritic cells and its potential synergy with toll-like receptor agonists. Viruses (MDPI). 2023, 15, 1198. doi.org/10.3390/v15051198.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2023
Citation:
Bian, Y., Walter, D., Zhang, C. 2023. Cytometric analysis authenticates adjuvanticity of interferon-? and suggests potential synergy with toll-like receptor agonists. J Immunol (2023) 210 (1_Supplement): 145.03. https://doi.org/10.4049/jimmunol.210.Supp.145.03
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2023
Citation:
Kristina Ly, Yuanzhi Bian, Riley DeHority, Debra Walter, XJ Meng, Chenming Zhang. Development of a Nanoparticle-Based Vaccine for PEDV. USDA National Institute of Food and Agriculture, A1511 Nanotechnology for Agriculture and Food Systems Annual meeting. Aug. 10-11, 2023. Knoxville, TN.
|
Progress 01/01/22 to 12/31/22
Outputs
Target Audience:No publications or presentations have resulted from the research effort in this reporting period. However, the news of the project (grant) was publicized by Virginia Tech, which could be seen by Virginia Tech personnel, alumni, and persons who follow Virginia Tech news.Virginia Tech researchers developing a new vaccine for a swine coronavirus VTx Virginia Tech Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?One graduate studentworked on this project together with an undergraduate researcher. Both of the students learned the basic molecular cloning techniques and became proficient with E. coli transformation for recombinant protein expression. The graduate student also gained valuable mentorship experience. How have the results been disseminated to communities of interest?
Nothing Reported
What do you plan to do during the next reporting period to accomplish the goals?The main focus of the next reporting period is to investigate how the spiked protein expressed in E. coli can be successfully recovered and purified. Based on our experiments, the protein is likely accumulated in the inclusion bodies of E. coli. Since the spike protein has a large domain embedded in the virus envelope (highly hydrophobic), we expect the solubilization of the protein may be challenging. We may explore to express the ectodomain of the protein, if the full length of the spike protein cannot be recovered and purified with high efficiency. If time permits, we will investigate how different molecular adjuvants can be included in the vaccine formulation.
Impacts
What was accomplished under these goals?
In this reporting period, we focused on progressing to accomplish the first objective. The first step of fabricating the vaccine is to express and purify the PEDV spike protein so it can be added to the surface of nanoparticles in preparation of the vaccine candidate. We have cloned the genes for the spike protein into E. coli, the system we are using to manufacture the spike protein. We have verified using PCR testing and DNA sequencing that the gene was cloned successfully. We have grown several batches of the E. coli with the spike protein gene and dosed them with a chemical called IPTG, to instruct the E. coli to produce the spike protein. We are in the process of developing a process to recover and purify the spike protein.
Publications
|
|