摘要
第一部分 CD28拮抗性类肽的筛选及其生物活性测定
目的
CD28-B7是T细胞活化最关键的，研究最清楚的协同刺激通路。阻断CD28-B7通路是治疗自身免疫病及防治移植排斥的重要策略之一。
目前阻断协同刺激通路的方法主要是用特异性单克隆抗体封闭，如用抗B7的单抗(抗-CD80／86)阻断CD28-B7通路，由于现有的单抗主要是鼠源性的，用于人体后可产生免疫反应，引起过敏反应和疗效的降低，而人单抗制备困难，基因工程产品技术要求和造价高，限制了其在临床的应用。因此，迫切需要研发出更为廉价、有效的阻断剂。
类肽是近年来发展起来的一种新型小分子受体或酶的阻断剂，可与受体的配体结合位点结合，阻断受体与天然配体的结合，起到受体抑制物的作用。类肽可根据受体或酶结合位点的三维空间结构设计并通过组合化学的方法合成。其优点是有高度的受体亲和力和选择性，在体内代谢稳定，制备简单，成本低。
本研究的目的是针对CD28与其配体结合的特异活性位点，用组合化学的方法合成CD28拮抗性类肽；利用噬菌体展示系统展示具有天然结构的CD28同源二聚体，以此建立一个简单、有效、高通量的体外筛选体系；以噬菌体展示CD28为靶标，筛选与CD28高亲和力结合的类肽药物；在体内外进一步检测CD28拮抗性类肽药物的生物活性，为CD28拮抗性类肽药物开发奠定基础。
方法
1．CD28拮抗性类肽设计、合成
根据CD28的三维空间结构及与B7结合关键基序，设计、合成CD28拮抗
Objective
The CD28-B7 pathway has been described as one of the best characterized, critical costimulatory pathway for T cell activation, so it may serve as a potential target for blocking T cell activation and thereby treating autoimmune diseases and ameliorating transplantation rejection.
The most common agent to block CD28-B7 costimulatory pathway is specific monoclonal antibodsy, such as anti-CD80/CD86mAbs. But monoclonal antibodies are mouse derived and have many side-effects in application. Otherwise, the production of humanized mAb is not only difficult but also expensive, so its application is limited. It is very necessary to make out more effective and cheaper blocking agents to block the costimulatory pathway.
Peptoid, oligomers of N-substituted glycines, is a new kind of blocking agent. It can interact with receptor or enzyme, so can be used as a small compound substitute for peptide to block the reaction of receptor with its natural ligand. Peptoid can be designed based on the receptor's conformation and synthesized by combinatory chemistry. Nowadays, peptoids attract more and more attention for their broad variety of biological activities and proteolytic stability.
The object of this research is to select peptoids that can block CD28-B7 pathway
引文
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