血型Ii单克隆抗体的研制及羊抗鼠二抗的制备

发布时间：2018-05-02 17:02

本文选题：Ii抗原 + IGnTc　；参考：《扬州大学》2006年硕士论文

【摘要】： Ii血型系统是第27个红细胞血型系统。它们没有自己的编码基因,i、I抗原分别由线性和分支型多聚N-乙酰半乳糖胺决定,因此其抗原决定簇为糖分子。新生儿脐血红细胞携带i抗原,随着生长发育,β-1,6-N-乙酰半乳糖胺转移酶活性的出现将2个N-乙酰基乳糖胺(N-AC)分别加到i抗原的线性多聚N-乙酰半乳糖胺结构的3和6位,从而使直链i抗原转变为分支型I抗原。本研究选用脐血红细胞做免疫原,用i(+)和i(-)红细胞进行筛选,得到三株抗i单克隆抗体。通过纯化i单克隆抗体(IgG)作为免疫原,获得了抗i的羊抗鼠二抗。抗i单克隆抗体及其二抗的研制为检测稀有血型ii,提供了必备的检测试剂。通过RT-PCR的方法成功地从胎盘组织中克隆了β-1,6-N-乙酰半乳糖胺转移酶基因(IgnTc),并构建了重组真核表达载体pcDNA3.1-IgnTc。重组质粒转染CHO细胞,使IgnTc在CHO细胞中的成功表达。IgnTc活性的出现将CHO细胞表面的i抗原转变为I抗原,获得了I阳性的CHO细胞株,为筛选抗I单克隆抗体及临床鉴定血清中相关I抗体的提供了重要的工具。本研究的实施将进一步确保输血安全,给输血患者带来福音,还将为在中国开展稀有血型的检测、普查工作,建立稀有血型库奠定基础。
[Abstract]:I blood group system is the 27 th red blood group system. They do not have their own coding gene, I. the antigens are determined by linear and branched polyN-acetylgalactosamine respectively, so their antigen determinants are sugar molecules. Umbilical cord blood red blood cells carried I antigen. With the development of growth and development, the activity of 尾 -1n 6-N-acetylgalactosamine transferase was added to the third and sixth position of linear polyN-acetylgalactosamine structure of I antigen, respectively. Thus the straight chain I antigen was transformed into branched I antigen. In this study, cord blood red blood cells were used as immunogen. The goat anti-mouse anti-I second antibody was obtained by purification of I monoclonal antibody (IgG) as immunogen. The development of anti-I monoclonal antibody and its second antibody provides a necessary reagent for the detection of rare blood group ii. The 尾 -1G 6-N-acetylgalactosamine transferase gene was cloned from placenta tissue by RT-PCR, and the recombinant eukaryotic expression vector pcDNA3.1-IgnTc. was constructed. The recombinant plasmid was transfected into CHO cells. The successful expression of IgnTc in CHO cells resulted in the transformation of I antigen from the surface of CHO cells to I antigen, and the I-positive CHO cell line was obtained. It provides an important tool for the screening of anti-I monoclonal antibodies and clinical identification of anti-I antibodies in serum. The implementation of this study will further ensure the safety of blood transfusion, bring good news to patients with blood transfusion, and lay the foundation for the testing and general investigation of rare blood groups in China and the establishment of a database of rare blood groups.
【学位授予单位】：扬州大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R392

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