微粒浓度荧光免疫分析法（PCFIA）：一种新型、快速且灵敏度高的免疫分析技术。

Particle concentration fluorescence immunoassay (PCFIA): a new, rapid immunoassay technique with high sensitivity.

作者信息

Jolley M E, Wang C H, Ekenberg S J, Zuelke M S, Kelso D M

出版信息

J Immunol Methods. 1984 Feb 24;67(1):21-35. doi: 10.1016/0022-1759(84)90082-6.

Abstract

A new solid-phase fluorescence immunoassay technique is described and is exemplified by the detection of murine monoclonal antibodies to human IgG in hybridoma culture supernatants and the detection of murine IgG. The assay is performed in a specially designed 96-well plate. For antibody detection, antigen bound to submicron polystyrene particles is bound to its specific antibody, which is in turn reacted with fluorescein-labeled affinity-purified goat anti-mouse IgG. The reaction is complete in 10 min at ambient temperature. The solid phase is separated from the reaction mixture by filtration, washed and the total particle-bound fluorescence is determined by front-surface fluorimetry. The sensitivity of the technique for antibody detection is equivalent to enzyme-linked immunoabsorbent assay and 2-4 ng/ml for murine IgG detection. It is readily amenable to automation.

摘要

本文描述了一种新的固相荧光免疫分析技术，并以检测杂交瘤培养上清液中抗人IgG的鼠单克隆抗体以及检测鼠IgG为例进行说明。该分析在专门设计的96孔板中进行。对于抗体检测，与亚微米聚苯乙烯颗粒结合的抗原与它的特异性抗体结合，然后该抗体再与荧光素标记的亲和纯化山羊抗鼠IgG反应。反应在室温下10分钟内完成。通过过滤将固相从反应混合物中分离出来，洗涤后，通过表面荧光法测定总颗粒结合荧光。该技术检测抗体的灵敏度与酶联免疫吸附测定相当，检测鼠IgG的灵敏度为2 - 4 ng/ml。它很容易实现自动化。