抗活化状态LM-1蛋白质单克隆抗体的制备及功能鉴定

发布时间：2018-11-17 12:10

【摘要】： LM-1蛋白质与白细胞的迁移和粘附密切相关,它表达在白细胞表面,通过与其配体结合,以介导白细胞从血液循环中进入肠道和中枢神经系统,LM-1的功能的异常可以导致肠道系统的慢性炎症和中枢神经系统损伤,从而引起一系列人类自身免疫疾病如asthma,arthritis,Crohn's disease,IBD和MS等。目前,众多国际上知名的医药公司正在开发以LM-1蛋白质为靶标的药物,目前唯一的一个以LM-1蛋白质为靶标的抗体药物会同时抑制活化和未活化的LM-1的功能,因此,该药物具有较大的副作用。我们的研究应用噬菌体展示技术,以活化状态的LM-1为抗原,筛选出了抗LM-1蛋白质的阳性噬菌体,纯化了表达于阳性噬菌体表面的scFV,用FACS和Flow Chamber系统鉴定后证明此scFV能特异性抗活化状态LM-1蛋白质并且具有非常高的效价。这就为进一步开发以活化状态的LM-1为靶标的抗体药物提供了基础,相比于上市的不能区分活化和未活化的LM-1蛋白质的抗体药物,我们的抗体具有明显的优势,由于能将药物靶向运输到膜表面存在活化状态LM-1的细胞,特异性对病变细胞进行杀伤,从而能保护正常的细胞,能显著的降低药物的副作用,具有非常好的应用前景。
[Abstract]:LM-1 protein is closely related to leukocyte migration and adhesion. It is expressed on the surface of white blood cells, and by binding with its ligand, it mediates leukocytes from blood circulation to the intestine and central nervous system. The abnormal function of LM-1 can lead to chronic inflammation of the intestinal system and damage of the central nervous system, resulting in a series of autoimmune diseases such as asthma,arthritis,Crohn's disease,IBD and MS. At present, many internationally renowned pharmaceutical companies are developing drugs targeting LM-1 protein. The only antibody drug targeting LM-1 protein will inhibit the function of activated and unactivated LM-1 at the same time. The drug has great side effects. Using phage display technique and activated LM-1 as antigen, we screened out the positive phage against LM-1 protein and purified scFV, expressed on the surface of positive phage. FACS and Flow Chamber analysis showed that the scFV could specifically resist the activated LM-1 protein and had a very high titer. This provides a basis for the further development of antibody drugs targeting activated LM-1, and our antibodies have obvious advantages over those that do not distinguish between activated and unactivated LM-1 proteins on the market. Because the drug can be targeted to the surface of the membrane there are activated LM-1 cells, the specific killing of pathological cells, thus can protect the normal cells, can significantly reduce the side effects of drugs, has a very good application prospects.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R392

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