1.利用噬菌体展示技术筛选Gal-α1，3-Gal结构的模拟肽 2.芋螺毒素M ⅦA的融合表达、纯化及其鉴定

发布时间：2018-01-08 13:10

本文关键词：1.利用噬菌体展示技术筛选Gal-α1，3-Gal结构的模拟肽 2.芋螺毒素M ⅦA的融合表达、纯化及其鉴定　出处：《浙江大学》2005年硕士论文　论文类型：学位论文

【摘要】：随着器官移植范围的扩大,器官紧缺问题日益紧迫。为了解决这一问题,异种器官的移植成为器官移植研究新的热点。在各种异种供体中,猪凭着自身器官形体、基因组成等特点成为许多研究者的首选。但是由于猪器官表面异种抗原Gal-α1,3-Gal能和人血清中的天然抗体发生结合,引发超急性排斥反应,随后激活补体的瀑布效应,导致内皮细胞死亡,使得异种器官难以在人体内存活。体内和体外的实验表明阻断人血清的天然抗体与猪器官表面抗原Gal-α1,3-Gal结合就可以防止超急性排斥反应,因此阻断人天然抗体与猪器官表面抗原Gal-α1,3-Gal结合便成为猪器官异种移植的关键。西非单叶豆凝集素(BS-I-B4)能结合α-1,3半乳糖(Gal-α1,3-Gal),因而可以用此凝集素模拟人天然抗体来进行α-1,3半乳糖(Gal-α1,3-Gal)模拟物的筛选。通过对西非单叶豆凝集素(BS-I-B4)在XCX15噬菌体随机文库中进行生物淘金筛选后,我们得到了序列为NCVSPYWCEPLAPSARA的小肽。经验证该小肽有竞争蜜二糖(具有Gal-α1,3-Gal结构?还是Gal-α1,6-Gal?)与凝集素BS-I-B4、A型人血清结合的能力,并可以用来抑制猪红细胞与凝集素BS-I-B4、抗B型血单克隆抗体及人的A型血血清(含人抗αGal的天然抗体)的凝集;证明该小肽可以模拟Gal-α1,3-Gal的结构,可以阻止人血清和猪器官发生的超急性排斥反应。
[Abstract]:With the expansion of the scope of organ transplantation, organ shortage is becoming more and more urgent. In order to solve this problem, xenotransplantation has become a new hotspot in organ transplantation. Pigs with their own organ form, gene composition and other characteristics have become the first choice for many researchers, but because the porcine organ surface xenogeneic antigen Gal- 伪 1 + 3-Gal can bind to the natural antibody in human serum. Hyperacute rejection is triggered, followed by the activation of the waterfall effect of the complement, leading to the death of endothelial cells. In vivo and in vitro experiments showed that blocking the binding of human serum natural antibodies to porcine organ surface antigen Gal- 伪 1 + 3-Gal could prevent hyperacute rejection. Therefore, blocking the binding of human natural antibody to porcine organ surface antigen Gal- 伪 1 + 3-Gal is the key to porcine organ xenotransplantation. The agglutinin BS-I-B4) can be used to simulate the human natural antibody to 伪 -1. It can be combined with galactose-galactose-3-galactose-3-galactose-3-galactose-galactose-3-galactose-galactose-3-galactosamine. (3) screening of galactose-Gal- 伪 1- (3-Gal3) mimics. BS-I-B4) was screened in XCX15 phage random library. We have obtained a small peptide with a sequence of NCVSPYWCEPLAPSARA. It has been proved that the small peptide has competitive honey disaccharide (Gal- 伪 1 + 3-Gal structure? Or Gal- 伪 1, 6-Galal? The ability to bind to human serum BS-I-B4A and to inhibit porcine erythrocyte and lectin BS-I-B4. Agglutination of monoclonal antibody against type B and human type A serum (natural antibody containing human anti-alpha Gal); It is proved that the small peptide can mimic the structure of Gal- 伪 1 + 3-Gal and prevent hyperacute rejection in human serum and pig organs.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R392.1

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