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

这一小型企业创新研究(SBIR)第一阶段项目的更广泛影响/商业潜力将是为全球民众提供安全、负担得起和可行的人类呼吸道合胞病毒(HRSV)可食用疫苗。呼肠孤病毒是一个关键的全球卫生问题，每年影响100万人，并导致超过16万人死亡，促使人们接种疫苗。天然食品衍生疫苗提供了一种安全、负担得起、非侵入性、不以鸡蛋为基础(降低过敏风险)、素食友好、有效和高效的抗原递送系统，因此比传统注射具有优势。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，这种疫苗可以无痛地在可咀嚼的口香糖状药丸配方中或在口服滴剂中给予。拟议的水果疫苗将通过可扩展的生产和分销将全球HRSV的发病率降至最低，提高全球可获得性和疫苗接种率。此外，它的一次性用品更少，更环保。该方法可广泛应用于其他口服给药平台。本项目一期工程将推动多肽口服给药平台的发展。该项目将提高产量和免疫原抗原(HRSV-F蛋白)在植物中的一致表达，无论是收获前和收获后，以及交付到体内之前和之后。该方法包括对HRSV-F蛋白的植物优化基因进行生物工程，通过阻断不必要的切割位点来提高其稳定性，同时保留HRSV-F免疫原所希望的保护性表位。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的樱桃西红柿将被收获，均质(泥)，然后冷冻干燥(冻干)，分别制成创新的疫苗泥或药丸。任务包括进行定性和定量分析，如颜色、浓度、pH、完整HRSV-F蛋白的浓度、其潜在降解产物和所需的表位/S，使用酶联免疫吸附试验(ELISA)、蛋白质印迹和高效液相色谱法进行分析。这些测试将有助于表征拟议的改变在新鲜水果、番茄泥和冷冻干燥水果丸中的HRSV-F蛋白表达、产量和稳定性方面的持久性和有效性。类似的评估也将在摄入后进行，在小鼠的肠道内容物和血液中。这一裁决反映了NSF的法定使命，并通过使用基金会的智力优势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.