CD28拮抗性类肽、CD137结合肽的筛选及生物活性测定

发布时间：2018-01-05 16:43

本文关键词：CD28拮抗性类肽、CD137结合肽的筛选及生物活性测定　出处：《山东大学》2006年博士论文　论文类型：学位论文

【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
[Abstract]:Screening and bioactivity determination of CD28 antagonistic peptides in part 1
objective
CD28-B7 is the most critical and most clear synergistic stimulus in T cell activation. Blocking CD28-B7 pathway is one of the most important strategies for autoimmune diseases and prevention of graft rejection.
At present, the main method of blocking the costimulatory pathway is closed with specific monoclonal antibodies, such as anti B7 monoclonal antibody (anti -CD80 / 86) blocking the CD28-B7 pathway, because existing monoclonal antibodies are mouse derived, for the human body can produce immune response, reduce allergic reaction and curative effect, and human monoclonal antibody system preparation of difficult technical requirements of genetic engineering products and high cost, which limit its clinical application. Therefore, the urgent need to develop a more inexpensive, effective blocking agent.
Is a kind of peptide inhibitor of novel small molecule receptor or enzyme developed in recent years, can be combined with the receptor ligand binding sites, blocking the receptor with its natural ligand, receptor inhibitor play role. The three-dimensional structure design according to the peptide receptor or enzyme binding sites and synthesized by combinatorial chemistry. Its advantage is high receptor affinity and selectivity in vivo metabolic stability, simple preparation and low cost.
The purpose of this study is specific to the active site of CD28 and its ligand binding, using the method of combinatorial chemistry synthesis of CD28 antagonistic peptides; displayed has a natural structure of the CD28 homodimer with two phage, in order to establish a simple, efficient, high-throughput screening system in vitro; show CD28 as a target in phage screening of peptide drugs has high affinity with CD28; further detection of CD28 antagonistic peptide drugs biological activity in vitro and in vivo, lay the foundation for the CD28 antagonistic peptide drug development.
Method
1.CD28 antagonistic peptide design, synthesis
According to the three-dimensional structure of CD28 and the key sequence of combining with B7, the design and synthesis of CD28 antagonism

【学位授予单位】：山东大学
【学位级别】：博士
【学位授予年份】：2006
【分类号】：R392

【共引文献】