biostudies-literatureCardenas MNIAID NIH HHS104155https://www.ebi.ac.uk/biostudies/studies/S-EPMC12593577biostudies-literatureUnknown6(4)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.STAR protocolsProtocol to measure the kinetics of MHC class II-influenza A binding using biolayer interferometry.PMC1259357775N93021C00014R01 AI165692Boons GJHunt AChopra PCardenas MRajao DSCompton SPerez DRfalseProtocol to measure the kinetics of MHC class II-influenza A binding using biolayer interferometry.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.2025-01-01T00:00:00Z2025 Oct2026-06-05T12:45:38.645Z2026-05-17T03:07:44.31ZS-EPMC125935774113818810.1016/j.xpro.2025.104155