抗利尿激素单克隆抗体的制备及初步应用

Preparation and Preliminary Application of Monoclonal Antibody against Human Antidiuretic Hormone

应用B淋巴细胞杂交瘤技术,成功地建立了分泌抗抗利尿激素(ADH)的单克隆抗体细胞株。新获的细胞株分泌的单克隆抗体类型属IgG2a。具诱生腹水抗体滴度可达1.2×10~4稀释度,灵敏度为2.5ng/ml。抗体的亲和常数为3.11×10~9L/mol,亲和力受温度影响明显。ADH McAb与催产素等11种神经肽进行竞争抑制试验均无交叉反应,静脉注射McAb可使新西兰白免尿量明显增加。用PAP免疫组化的方法对人和大鼠下丘脑神经细胞进行ADH定位检测,视上核及室旁核的神经细胞及神经纤维均有明显清晰的染色颗粒。应用放射免疫方法检测大鼠垂体,下丘脑组织ADH含量分别为186.46±53.23ng/mg,3.32±1.69ng/mg。因此,抗ADH McAb在八DH的组织学定位,组织含量的测定,以及作为中和剂应用在各种疾病的研究上都有重要的意义。

A hybridoma secreting monoclonal antibody (McAb) against human antidiu-retic hormone (ADH) was obtained by fusion of Sp2/0 myeloma cells with spleen cells from BALB/c mice immunized with ADH. The McAb was identified as IgG2a. Radio-immunoassay (RIA) indicated that the liter and affinity constant for this antibody was 1.2×104 and 3×109 L/mol, respectively. The affinity was affected markedly by temperature. Competitive inhibitory test demonstrated that McAb against ADH showed no cross reactivity with eleven other types of neuropeptides. 'The volume of urine increased significantly after intravenous injection of McAb in NewZealarid rabbits. Immunoh-istological examination using PAP showed that ADH was localized in the hypothalamic nervous nuclei in human and rat with strong positive staining in the suprapptical nuclei and paraventricular nuclei. The amount of ADH in the pituitary gland and hypothala-mic tissues as measured by RIA was 186.46±53.23ng/mg and 3.32±1.69ng/mg, respectively. These findings suggested that this McAb against ADH is of great importance in the histological localization of ADH, measurement of ADH in tissues and application of the antibody as neutralizer in some diseases.