Cell-based flow cytometry assay for simultaneous detection of multiple autoantibodies in a single serum sample.

Autor: Tanimura Y; Cellular and Structural Physiology Institute (CeSPI), Nagoya University, Nagoya, 464-8601, Japan. Electronic address: yukihiro.cesp@tmd.ac.jp., Hiroaki Y; Cellular and Structural Physiology Institute (CeSPI), Nagoya University, Nagoya, 464-8601, Japan., Mori M; Department of Neurology, Graduate School of Medicine, Chiba University, Chiba, 260-8670, Japan., Fujiyoshi Y; Cellular and Structural Physiology Institute (CeSPI), Nagoya University, Nagoya, 464-8601, Japan; CeSPIA Inc., Tokyo, 100-0004, Japan. Electronic address: yoshi.cesp@tmd.ac.jp.
Jazyk: angličtina
Zdroj: Analytical biochemistry [Anal Biochem] 2022 Aug 01; Vol. 650, pp. 114721. Date of Electronic Publication: 2022 May 13.
DOI: 10.1016/j.ab.2022.114721
Abstrakt: Accurate serologic evaluation of autoantibodies in patients with autoimmune diseases is critical. In the present study, we established a live cell-based assay for simultaneous detection of multiple autoantibodies in a single serum sample. Autoantibody seropositivity was determined by 3-color flow cytometry using live Chinese hamster ovary cells transiently expressing a target protein of interest fused to enhanced green fluorescent protein and labeled with Alexa Fluor 647 and Hoechst 33342. As a representative example, we applied the strategy for simultaneous detection of 2 recently established biomarkers for central nervous system autoimmune inflammatory demyelinating disorders, anti-aquaporin-4 autoantibody and anti-myelin oligodendrocyte glycoprotein autoantibody, in a single serum sample. This analysis revealed the coexistence of these 2 autoantibodies. We demonstrated that this assay can simultaneously detect 3 different autoantibodies. We propose a quadrant gating strategy of flow cytometry contour plots to clearly distinguish seropositive sera from seronegative sera regardless of the extent of the background signal level or the autoantibody titer. This novel and practical method using a combination of fluorescent proteins and fluorochromes to simultaneously detect multiple autoantibodies improves the efficiency of evaluating serum samples, and therefore provides significant benefits to both the patient and the healthcare professionals performing autoantibody testing.
(Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
Externí odkaz: