Work

Protocol to measure the kinetics of MHC class II-influenza A binding using biolayer interferometry

Year: 2025

Type: article

Abstract: Recent work has shown that major histocompatibility complex (MHC) class II is a functional receptor for some influenza A virus (FLUAV) subtypes. Here, we present a protocol to measure the kinetics of MHC class II-influenza A binding using biolayer interferometry. We describe steps for producing recombinant human leukocyte antigen DR isotype (HLA-DR) MHC class II using a baculovirus expression system. The HLA-DR antigen is then modified for its utilization as a biolayer interferometry (BLI) probe for kinetic experiments using whole FLUAV viruses. This protocol allows for the screening of different FLUAV subtypes and their affinity for MHC class II. • Steps for production of dimeric MHC class II proteins using a baculovirus expression system • Procedure to screen for influenza A viruses and MHC class II binding using BLI • Instructions to quantify affinity of whole influenza A viruses for MHCII Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Recent work has shown that major histocompatibility complex (MHC) class II is a functional receptor for some influenza A virus (FLUAV) subtypes. Here, we present a protocol to measure the kinetics of MHC class II-influenza A binding using biolayer interferometry. We describe steps for producing recombinant human leukocyte antigen DR isotype (HLA-DR) MHC class II using a baculovirus expression system. The HLA-DR antigen is then modified for its utilization as a biolayer interferometry (BLI) probe for kinetic experiments using whole FLUAV viruses. This protocol allows for the screening of different FLUAV subtypes and their affinity for MHC class II.

Source: STAR Protocols

DOI: 10.1016/j.xpro.2025.104155