Faresjö Maria
School of Health Sciences, Department of Natural Science and Biomedicine, The Biomedical Platform, Jönköping University, SE-551 11, Jönköping, Sweden,
Methods Mol Biol. 2014;1172:87-96. doi: 10.1007/978-1-4939-0928-5_7.
The fluorochrome Luminex technique is a bead-based sandwich immunoassay that combines the enzyme-linked immuno sorbent assay (ELISA) with flow cytometry. The Luminex technique allows multiple cytokines to be measured simultaneously in small volumes and provides a convenient and sensitive tool for the detection of a large number of, e.g., extracellular secreted cytokines to characterize cytokine profiles.The technique is based on the so-called microspheres (beads) that serve as a solid phase for molecular detection. These individually dyed micro-beads have monoclonal antibodies directed against the cytokines and chemokines of interest and allow simultaneous detection of up to nearly 100 cytokines and chemokines in a dual-laser flow analyzer. Immune markers can be detected in serum and plasma samples as well as in cell culture supernatants from in vitro-stimulated peripheral blood mononuclear cells (PBMC).This chapter describes the Luminex technique for detection of multiple cytokines by magnetic bead sandwich immunoassay, with a special focus on some important pre-analytical factors, such as cell separation, cryopreservation, and PBMC thawing that may affect the detection outcome of immune markers. This method can also be easily adapted to measuring other biomarkers in biological samples.
荧光染料Luminex技术是一种基于微珠的夹心免疫测定法，它将酶联免疫吸附测定法（ELISA）与流式细胞术相结合。Luminex技术能够在小体积样本中同时检测多种细胞因子，为检测大量细胞因子（如细胞外分泌的细胞因子以描绘细胞因子谱）提供了一种便捷且灵敏的工具。该技术基于所谓的微球（珠子），这些微球作为分子检测的固相。这些经过单独染色的微珠带有针对目标细胞因子和趋化因子的单克隆抗体，在双激光流式分析仪中能够同时检测多达近100种细胞因子和趋化因子。免疫标志物可在血清和血浆样本以及体外刺激的外周血单个核细胞（PBMC）的细胞培养上清液中检测到。本章介绍了通过磁珠夹心免疫测定法检测多种细胞因子的Luminex技术，特别关注一些重要的分析前因素，如细胞分离、冷冻保存以及PBMC解冻，这些因素可能会影响免疫标志物的检测结果。该方法也可轻松适用于测量生物样本中的其他生物标志物。
