摘要
本文运用淋巴细胞杂交瘤技术获得5个产生抗碱性磷酸酶单克隆抗体的杂交瘤细胞株,同时设计一种改良ELISA方法用于筛选抗碱性磷酸酶单克隆抗体。筛选出的单克隆抗体能与碱性磷酸酶结合形成可溶性的复合物,即碱性磷酸酶一抗碱性磷酸酶抗体(APAAP)复合物。这些APAAP复合物仍保留有较高的酶活性,并可改良ELISA方法确定其酶一抗酶抗体的配制比例。在免疫组化染色中,APAAP复合物通过桥联二抗及显色系统能在组织原位较好地显示与一抗相对应的抗原物质。这种APAAP桥联酶标记技术显色鲜明,染色背景清晰,染色后的组织形态仍保存完好。APAAP桥联酶标记技术结合免疫金银染色进行免疫组化双重标记也取得了满意的标记效果。初步应用结果表明APAAP桥联酶标记技术是一种简便、灵敏、易于推广的实用免疫组织化学方法。
Mouse monoclonal antibodies to alkaline phosphatase have been prepared with hybrid-ma technique. A specific assay in which the alkaline phosphatase-anti-alkaline phosphatase (APAAP) complex was captured by a anti-mouse Ig antibody was used for sceening hibrid cells. Five cell clones producing antibodies forming active soluble APAAP complexes with phosphatase were selected by this assay. The APAAP complexes was used in an unlabeled antibody bridge technique for immunostaining. This procedure-the APAAP method——gives excellent immunohistochemical labeling of tissue sections and cell smears in clarity and intensity. Surface or cytoplasmic determinants can be stained with the APAAP method. The APAAP technique could be used for doube immunohistochemical labeling combined with immuno-gold silver staining. The result indicates
that APAAP technique is a reliable sensitive and practical method.
出处
《免疫学杂志》
CAS
CSCD
北大核心
1989年第3期192-195,共4页
Immunological Journal
关键词
单克隆抗体
APAAP复合物
Monoclonal antibodies,APAAP complex,Imtnunoenzymatic labeling
