Li Ting, Wang Wenyan, Cheng Yingying, Han Wenling
Department of Immunology, School of Basic Medical Sciences, Peking University Health Science Center, Key Laboratory of Medical Immunology, Ministry of Health (Peking University), Beijing, 100191, China; Peking University Center for Human Disease Genomics, Beijing, 100191, China.
Department of Immunology, School of Basic Medical Sciences, Peking University Health Science Center, Key Laboratory of Medical Immunology, Ministry of Health (Peking University), Beijing, 100191, China; Peking University Center for Human Disease Genomics, Beijing, 100191, China; Tsinghua University School of Medicine, Beijing 100084, China.
J Biosci Bioeng. 2018 Mar;125(3):359-364. doi: 10.1016/j.jbiosc.2017.10.008. Epub 2017 Nov 20.
FAM19A4 (family with sequence similarity 19 member A4; also TAFA4) is a classical secretory protein expressed mainly in the central nervous system and upregulated significantly in lipopolysaccharide (LPS)-stimulated monocytes and macrophages. It is a novel cytokine ligand of formyl peptide receptor 1 (FPR1), showing chemotactic activities on macrophages and promoting the phagocytosis capacity and the release of reactive oxygen species (ROS) by macrophages upon zymosan stimulation. Based on the same detection principle as enzyme-linked immunosorbent assay (ELISA), we developed a sandwich immunoassay for quantitative detection of FAM19A4 in biological fluids by flow cytometry, with latex beads as solid carrier. The method showed good performance in a wide range of 39-10,000 pg/mL and possessed excellent specificity, good precision, and favorable recovery in several different matrices. Native FAM19A4 secreted by phorblo 12-myristate 13-acetate (PMA) and LPS stimulated THP-1 cells could also be detected by this method. This method will be much helpful to FAM19A4 studies.
FAM19A4（序列相似性家族19成员A4；也称为TAFA4）是一种典型的分泌蛋白，主要在中枢神经系统中表达，在脂多糖（LPS）刺激的单核细胞和巨噬细胞中显著上调。它是甲酰肽受体1（FPR1）的一种新型细胞因子配体，对巨噬细胞具有趋化活性，并在酵母聚糖刺激下促进巨噬细胞的吞噬能力和活性氧（ROS）释放。基于与酶联免疫吸附测定（ELISA）相同的检测原理，我们开发了一种以乳胶珠为固相载体的流式细胞术夹心免疫测定法，用于定量检测生物体液中的FAM19A4。该方法在39-10000 pg/mL的宽范围内表现良好，在几种不同基质中具有优异的特异性、良好的精密度和良好的回收率。该方法还可检测佛波醇12-肉豆蔻酸酯13-乙酸酯（PMA）和LPS刺激的THP-1细胞分泌的天然FAM19A4。该方法将对FAM19A4的研究有很大帮助。
