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

这项小企业创新研究（SBIR）第一阶段项目的更广泛影响/商业潜力将是为全球人口提供一种安全、负担得起和可行的抗人类呼吸道合胞病毒（hRSV）的食用疫苗。hRSV是一个严重的全球健康问题，每年影响6400万人，造成16万多人死亡，这促使人们开发疫苗。天然食物来源的疫苗提供了一种安全、负担得起、非侵入性、非蛋基（降低过敏风险）、素食友好、有效和高效的抗原递送系统，比传统注射更具优势。该项目通过开发一种樱桃番茄为基础的口服疫苗来满足这一需求，这种疫苗可以无痛地以可咀嚼的胶状药丸制剂或口服滴剂的形式进行注射。拟议的水果疫苗将通过大规模生产和分销，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，使用更少的一次性用品更环保。这种方法可以广泛应用于其他口服给药平台。该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，以及输送到体内前后。该方法涉及对hrv - f蛋白的植物优化基因进行生物工程，通过阻断不需要的切割位点来提高其稳定性，同时保留hrv - f免疫原所需的保护性表位。在番茄果实中表达稳定的hRSV-F的转基因植株将在温室中种植。含有hrsv - f的樱桃番茄将被收获、均质（泥），然后冻干（冷冻干燥），分别制成创新的疫苗泥或药片。任务包括进行定性和定量分析，如完整hRSV-F蛋白的颜色、一致性、pH、浓度、其潜在降解产物和所需表位/s，并使用酶联免疫吸附试验（ELISA）、western-blot和高效液相色谱分析。这些试验将有助于表征对新鲜水果、番茄泥和冻干水果丸中hRSV-F蛋白表达、产量和稳定性的拟议变化的持久性和有效性。同样的评估也将在小鼠摄入后的肠道内容物和血液中进行。该奖项反映了美国国家科学基金会的法定使命，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。

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.