用固相间接免疫荧光分析法检测杂交瘤细胞在培养物中合成的单克隆流感抗体。

Detection of monoclonal influenza antibodies synthesized in culture by hybridoma cells with a solid-phase indirect immunofluorometric assay.

作者信息

Phillips D J, Kendal A P, Webster R G, Feorino P M, Reimer C B

出版信息

J Virol Methods. 1980;1(5):275-83. doi: 10.1016/0166-0934(80)90024-5.

Abstract

A solid-phase indirect immunofluorometric assay for measuring reactions of mouse monoclonal antibodies with antigen has been developed, with influenza virus as a model. Purified IgG from hyperimmune rabbit sera is covalently linked to polyaminostyrene beads, to which influenza viruses are then bound immunologically to make solid-phase antigens. Alternatively, the virus is covalently coupled directly to the beads. Mouse antibodies, produced by hybridoma cells in culture, are reacted with constant amounts of solid-phase antigens, and then indirectly quantitated by adding FITC-labeled antimouse Ig and measuring the fluorescent intensity with a filter-fluorometer. The assay system permits rapid screening for low levels of antibodies synthesized by hybridoma cells in culture. It is about 25- to 150-fold more sensitive than hemagglutination inhibition tests in detecting monoclonal antibodies reactive with influenza virion HA protein.

摘要

已开发出一种用于检测小鼠单克隆抗体与抗原反应的固相间接免疫荧光测定法，以流感病毒为模型。从超免疫兔血清中纯化的IgG与聚氨基聚苯乙烯珠共价连接，然后通过免疫方式将流感病毒结合到这些珠子上以制成固相抗原。或者，病毒直接与珠子共价偶联。培养的杂交瘤细胞产生的小鼠抗体与恒定数量的固相抗原反应，然后通过添加异硫氰酸荧光素（FITC）标记的抗小鼠Ig并使用滤光荧光计测量荧光强度进行间接定量。该测定系统可快速筛选培养的杂交瘤细胞合成的低水平抗体。在检测与流感病毒粒子HA蛋白反应的单克隆抗体方面，它比血凝抑制试验灵敏约25至150倍。