抗人粒细胞单克隆抗体的制备及其鉴定

Preparation and Characterization of Monoclonal Antibody Against Human Granulocytes

目的 制备抗人粒细胞单克隆抗体 (McAb) ,并对其组织特异性及相关特性进行鉴定。方法 采用分离的人全白细胞免疫Balb/c小鼠 ,按常规融合方法并经ELISA筛选后进行McAb特性研究。结果 成功制备 1株抗人粒细胞McAb ,命名为SZ - 10 2。ELISA法测定其腹水效价为 5× 10 -5。琼脂糖双扩散鉴定均为IgG1类。流式细胞仪及SP免疫组化法测定表明SZ - 10 2McAb与外周血液中的粒细胞、单核细胞呈强阳性反应 ,与淋巴细胞、红细胞、血小板不反应 ;SZ - 10 2抗原在正常人肝、肺、胸腺、脾、淋巴结组织中也表达。免疫沉淀测定抗原分子量为 2 0kD左右的蛋白多肽。结论 抗人粒细胞McAb的研制将有助于临床肿瘤感染病灶和隐匿性炎症的放免显像诊断研究。

Objective To prepare monoclonal antibody (McAb) against human granulocytes and determine the characterizations of McAb(SZ-102).Methods Balb/c mouse was immunized by intraperitional route. After hybridizing and screening, and McAb's characterizations were determined.Results McAb(SZ-102) against human granulocytes was obtained . McAb(SZ-102) in ascite fluid was 5×10 -5 . The antibody belongs to IgG 1 subclass. It was confirmed by immunohistochemical methods that SZ-102 reacted selectively with human granulocyte,monocytes, and with their bone marrow precursors, while human lymphocytes and red blood cells remained negative.SZ-102 antigen was also expressed in liver, lung, thymus gland, spleen, lymph node. Immunoprecipitation demonstrated that molecular weight of polypeptide was 20kD under reducing condition.Conclusion McAb(SZ-102) against human granulocytes would provide us an effective and reliable diagnostic method in detecting inflammational areas of carcinomas and clinically concealed infectious sites.