SBIR Phase I: Stabilization of the desired epitopes of hRSV-F protein for efficient absorption through the gut

SBIR 第一阶段：稳定 hRSV-F 蛋白所需表位，以实现肠道有效吸收

• 批准号: 2026281
• 负责人: Swarnamali Rupassara
• 金额: $ 25.6万
• 依托单位: FruitVaccine Incorporation
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-12-01 至 2021-11-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2026281&HistoricalAwards=false
• 关键词: SBIR; Phase; Stabilization; desired; epitopes

The broader impact/commercial potential of this Small Business Innovation Research (SBIR) Phase I project will be to provide global populations with a safe, affordable and viable edible vaccine against the human respiratory syncytial virus (hRSV). hRSV is a critical global health problem, annually affecting 64 million people and causing over 160,000 death, motivating a vaccine. Natural food-derived vaccines offer advantages over traditional injections by providing a safe, affordable, non-invasive, non-egg-based (reduced allergenic risk), vegan-friendly, effective and efficient antigen-delivery system. The project addresses this need by developing a cherry-tomato-based oral vaccine that can be administered painlessly in a chewable gummy-like pill formulation or in oral drops. The proposed fruit-vaccines will minimize the global incidence of hRSV through scalable production and distribution, increasing global accessibility and vaccination rates. Furthermore, with fewer disposables it is more environmentally friendly. This method could apply broadly to other oral-delivery platforms.This Phase I project will advance peptide-based oral-delivery platforms. The project will improve the yield and consistent expression of the immunogenic antigen (hRSV-F protein) in the plant, both pre- and post-harvest, as well as before and after delivery into the body. The approach involves bioengineering the plant-optimized gene for the hRSV-F protein to improve its stability by blocking undesirable cleavage sites while retaining the desirable protective epitopes of the hRSV-F immunogen. Transgenic plants expressing the stabilized hRSV-F in the tomato fruits will be grown in a greenhouse. The hRSV-F-containing cherry-tomatoes will be harvested, homogenized (pureed), and then lyophilized (freeze-dried) to formulate innovative vaccine purees or pills, respectively. Tasks include conducting qualitative and quantitative analyses, such as color, consistency, pH, concentration of intact hRSV-F protein, its potential degradation products and desired epitope/s, analyzed using enzyme-linked immunosorbent assay (ELISA), western-blot and high-performance liquid chromatography. These tests will help characterize the persistence and effectiveness of the proposed changes on hRSV-F protein expression, yield, and stability in fresh-fruits, tomato-puree and in the freeze-dried fruit-pills. Similar assessments will be performed also post-ingestion, in the gut contents and in blood of mice.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

这项小型企业创新研究（SBIR）第一阶段项目的更广泛影响/商业潜力将为全球人口提供一种安全、负担得起和可行的抗人呼吸道合胞病毒（hRSV）的可食用疫苗。hRSV是一个严重的全球健康问题，每年影响6400万人并导致超过160，000人死亡，促使疫苗。天然食物来源的疫苗通过提供安全、负担得起、非侵入性、非基于鸡蛋（降低过敏风险）、素食友好、有效和高效的抗原递送系统而提供优于传统注射的优势。该项目通过开发一种以樱桃番茄为基础的口服疫苗来满足这一需求，该疫苗可以以咀嚼胶样药丸或口服滴剂的形式无痛地施用。 拟议的水果疫苗将通过可规模化的生产和分销，提高全球可及性和疫苗接种率，最大限度地减少hRSV的全球发病率。此外，一次性用品更少，更环保。该方法可广泛应用于其他口服给药平台，本一期项目将推进基于肽的口服给药平台。 该项目将提高植物中免疫原性抗原（hRSV-F蛋白）的产量和一致表达，包括收获前和收获后，以及交付到体内之前和之后。该方法涉及对hRSV-F蛋白的植物优化基因进行生物工程改造，以通过阻断不需要的切割位点来提高其稳定性，同时保留hRSV-F免疫原的理想保护性表位。在番茄果实中表达稳定的hRSV-F的转基因植物将在温室中种植。将收获含hRSV-F的樱桃番茄，均质化（泥化），然后冻干（冷冻干燥）以分别配制创新的疫苗泥或丸剂。任务包括进行定性和定量分析，例如使用酶联免疫吸附试验（ELISA）、蛋白质印迹和高效液相色谱法分析完整hRSV-F蛋白的颜色、稠度、pH值、浓度、其潜在降解产物和所需表位。 这些测试将有助于表征所提出的变化对新鲜水果、番茄泥和冻干水果丸中的hRSV-F蛋白表达、产量和稳定性的持续性和有效性。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。