抗ICAM-1单链抗体的制备及其生物学活性研究

发布时间：2018-05-02 20:41

本文选题：ICAM-1 + 单链抗体　；参考：《吉林大学》2008年博士论文

【摘要】： 炎症是机体的一种重要的基本病理过程。各种炎症虽然有其特殊性,但从局部的病理变化看来,它们有着不少共同点,即都是机体对致炎因子的损伤作用所产生的一种反应,最新的医学研究发现,当炎症的程度严重或发展成慢性炎症时,它可能导致肿瘤、心血管疾病、糖尿病、自身免疫性疾病以及与年龄相关的疾病(如关节炎、早老性痴呆病、骨质疏松症等)的发生。细胞间粘附分子-1(ICAM-1)作为重要的致炎因子,在炎症反应、免疫监督等过程中起着重要的作用。以上许多疾病过程中可见ICAM-1的异常表达,因此阻遏ICAM-1的表达为以上疾病的诊断和治疗提供了新的策略。本研究在已制备的抗ICAM-1单克隆抗体的基础上,从分泌该单抗的杂交瘤细胞株中克隆出VH和VL基因,组装成ScFv基因,将ScFv基因插入原核表达载体pET-22b(+)中,转化宿主菌中进行表达,之后对表达的包涵体进行了复性和纯化。最后通过体外细胞实验和体内动物实验检测ScFv抗体的生物学活性。细胞实验表明,制备的ScFv能有效地抑制内皮细胞与单核细胞之间的粘附;动物实验证明此ScFv对小鼠无菌性炎症有明显的抑制作用。本课题为进一步治疗炎症性疾病奠定了基础。
[Abstract]:Inflammation is an important basic pathological process of the body. Although various kinds of inflammation have their own particularity, but from the local pathological changes, they have a lot in common, that is, they are all a kind of reaction of the body to the injury of inflammatory factor, the latest medical research found, When inflammation is severe or develops into chronic inflammation, it can lead to tumours, cardiovascular diseases, diabetes, autoimmune diseases, and age-related diseases such as arthritis, Alzheimer's disease, osteoporosis, etc. Intercellular adhesion molecule-1 (ICAM-1), as an important inflammatory factor, plays an important role in inflammatory response and immune supervision. The abnormal expression of ICAM-1 can be seen in many of the above diseases, so the repressive expression of ICAM-1 provides a new strategy for the diagnosis and treatment of these diseases. In this study, VH and VL genes were cloned from hybridoma cell lines secreting the monoclonal antibodies against ICAM-1, and ScFv gene was assembled into the prokaryotic expression vector pET-22b (). The VH and VL genes were expressed in the transformed host strain. After that, the expressed inclusion bodies were renatured and purified. Finally, the biological activity of ScFv antibody was detected by in vitro cell experiment and in vivo animal experiment. Cell experiments showed that the prepared ScFv could effectively inhibit the adhesion between endothelial cells and monocytes, and that the ScFv could inhibit the aseptic inflammation in mice. This topic has laid the foundation for the further treatment of inflammatory diseases.
【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2008
【分类号】：R392.12

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