Development of VesiVax Carbohydrate Conjugation Chemistry

描述：针对PA-10-150，我们将测试这一假设，即VesiVax(R)结合佐剂脂囊(CALV)平台可以提供一种有效的方法来开发碳水化合物结合疫苗，该方法可刺激对非结合碳水化合物抗原的卓越免疫反应，并提供比目前使用的结合疫苗技术更大的灵活性来控制对碳水化合物抗原的免疫反应。为了证明VesiVax(R)CALV平台的实用性，我们建议开发一种多糖结合技术，其商业化目标是对VesiVax(R)CALV系列产品进行易于使用的改进。预计该项目的成功实施将提供一种新的工具，使免疫学家和疫苗专家能够评估他们感兴趣的碳水化合物抗原。多糖正在成为包括病毒、细菌、真菌和癌症在内的各种病原体的有希望的疫苗靶点。在此SBIR第一阶段应用中，我们将开发一种简单的碳水化合物偶联方法，将多糖抗原偶联到蛋白质支架或含有脂胺的脂质体上的游离胺。具体地说，使用肠沙门氏菌(伤寒)的胶囊多糖Vi作为我们的模型碳水化合物抗原，我们将Vi直接偶联到含有Toll样受体4(TLR4)激动剂的VesiVax(R)CALV，或连接到基于白喉毒素突变CRM197的蛋白质支架，该突变被认为具有C-末端半胱氨酸残基(CRM197-Cys)。然后，Vi-CRM197-Cys将与VesiVax(R)CALV结合。然后，含有TLR4激动剂的VesiVax(R)VCalv结合物将在小鼠免疫模型中进行测试，以证明所提出的方法的实用性。

DESCRIPTION: In response to PA-10-150, we will test the hypothesis that the VesiVax(R) Conjugatable Adjuvant Lipid Vesicle (CALV) platform can provide an efficient method to develop carbohydrate conjugate vaccines that stimulates superior immune responses to unconjugated carbohydrate antigens and also provides more flexibility to control the immune response to carbohydrate antigens over currently employed conjugate vaccine technologies. To demonstrate the utility of the VesiVax(R) CALV platform for this purpose, we propose to develop a polysaccharide conjugation technology with the commercialization goal of producing an easy-to-use modification to the VesiVax(R) CALV line of products. It is anticipated that successful execution of this project will provide a new tool that will enable immunologists and vaccinologists to evaluate their carbohydrate antigens of interest. Polysaccharides are emerging as promising vaccine targets for a variety of pathogens including viruses, bacteria, fungi, and cancers. In this SBIR Phase I application, we will develop a simple carbohydrate conjugation method that couples polysaccharide antigens to free amines on either a protein scaffold or liposomes containing lipid amines. Specifically, using the capsular polysaccharide Vi from Salmonella enterica (typhi) as our model carbohydrate antigen, we will couple Vi directly to VesiVax(R) CALVs containing a Toll-like Receptor 4 (TLR4) agonist or to a protein scaffold based on the mutant of diphtheria toxin known as CRM197 engineered to have a C-terminal cysteine residue (CRM197-Cys). The Vi-CRM197-Cys will then be conjugated to the VesiVax(R) CALVs. The VesiVax(R) Vi CALV conjugates containing the TLR4 agonist will then be tested in a mouse model of immunization to demonstrate the utility of the proposed approach.