用合成多肽免疫制备人α-半乳糖苷酶A单克隆抗体  

Preparation of Anti-Human α-Galactosidase A Monoclonal Antibody with Synthetic Peptide

作　　者：徐莉娟[1] 季守平[1] 高红伟[1] 李素波[1] 刘至玄[1] 田曙光[1] 宫锋[1] 章扬培[1] 

机构地区：[1]军事医学科学院野战输血研究所,北京100850

出　　处：《生物技术通讯》2007年第4期600-603,共4页Letters in Biotechnology

基　　金：国家重点基础研究发展规划项目(2002CB713804)

摘　　要：目的:获得效价高的、特异性强的抗人α-半乳糖苷酶A(α-GalA)单克隆抗体,用于法布莱氏病的诊断。方法:分析并合成人α-GalA的优势抗原表位,合成多肽,将其与载体匙孔虫戚血蓝蛋白偶联,免疫小鼠,采用甲基纤维素半固体培养基筛选杂交瘤细胞株;用酶联免疫、SDS-PAGE、Western印迹和染色体核型分析等方法对杂交瘤细胞及其产生的单抗进行鉴定。结果与结论:筛选到1株杂交瘤细胞株,获得了高效价、特异性强、亲和力高的抗人α-GalA的单克隆抗体,抗体的亚型为IgG1亚型。Objective： To obtain high titer and high specific human α-alactosidase A（α-GalA） monoclonal antibody for Fabry disease diagnosis. Methods： Dominant epitope of human α-GalA was analyzed and synthesized. The synthesized peptides were conjugated to KLH and BALB/c mice were immunized with conjugated peptides. The hybridoma cell lines were selected with CM semisolid culture medium containing HAT. The hybridoma cell lines and their monoclonal antibodies were identified by enzyme linked immunosorbent assay（ELISA）, SDS-PAGE, Western-blot as well as chromosome analysis. Results ＆ Conclusion： One hybridoma cell line producing high titer, high specific and high affinity anti human or- GalA monoclonal antibody was obtained. The antibody was identified to be IgG1 subtype.

关 键 词：Α-半乳糖苷酶A 单克隆抗体 合成多肽 抗原表位 半固体培养基 

分 类 号：R392[医药卫生—免疫学] Q789[医药卫生—基础医学]