MilliporeSigma

Detection of Neutralizing Anti-Type 1 Interferon Autoantibodies.

Current protocols (2022-08-18)
Elana R Shaw, Lindsey B Rosen, Li Ding, Steven M Holland, Helen C Su
ABSTRACT

Autoantibodies (autoAbs) that neutralize type 1 interferons (T1IFNs) are a major risk factor associated with developing critical COVID-19 disease and are most commonly found in individuals over age 70 and in patients with genetic or acquired thymic defects. Swift identification of autoAb-positive individuals may allow targeted interventions to prevent critical COVID-19 disease. Herein, we provide a workflow and protocols aimed at rapidly identifying individuals who are autoAb positive from a large cohort. Basic Protocol 1 describes a multiplex particle-based assay to screen large cohorts of individuals for binding levels of anti-T1IFN autoAbs, and Basic Protocol 2 describes a functional assay to test if autoAbs in patient plasma can block T1IFN-induced JAK/STAT signaling. © Published 2022. This article is a U.S. Government work and is in the public domain in the USA. Basic Protocol 1: Multiplex particle-based bead assay to screen for binding levels of anti-type 1 interferon autoantibodies Alternate Protocol: Multiplex particle-based bead assay to screen for binding levels of anti-type 1 interferon immunoglobulin subtypes and isotypes Support Protocol: Coupling type 1 interferons (IFN-α, IFN-β, and IFN-ω) to magnetic beads Basic Protocol 2: pSTAT1 functional assay to test for neutralization activity of anti-type 1 interferon autoantibodies.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Anti-Human IgG3−Biotin antibody, Mouse monoclonal, clone HP-6050, purified from hybridoma cell culture
Sigma-Aldrich
Anti-Human IgG1−Biotin antibody, Mouse monoclonal, clone 8c/6-39, purified from hybridoma cell culture
Sigma-Aldrich
Anti-Human IgG2−Biotin antibody, Mouse monoclonal, clone HP-6014, purified from hybridoma cell culture
Sigma-Aldrich
Anti-Human IgG4−Biotin antibody, Mouse monoclonal, clone HP-6025, purified from hybridoma cell culture