Protocol 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.
Authors
- Geert‐Jan Boons (ORCID: https://orcid.org/0000-0003-3111-5954)
- Pradeep Chopra (ORCID: https://orcid.org/0000-0002-6003-4574)
- Ashkon Hunt
- Susan R Compton (ORCID: https://orcid.org/0000-0001-9644-1561)
- Daniel R. Pérez (ORCID: https://orcid.org/0000-0002-6569-5689)
- Matías Cárdenas (ORCID: https://orcid.org/0000-0001-8247-3669)
- Daniela S. Rajão (ORCID: https://orcid.org/0000-0002-0772-0065)
Institutions
- University of Georgia (US)
- Utrecht University (NL)
Publication Details
- Journal
- STAR Protocols
- Published
- 2025-10-25
- DOI
- https://doi.org/10.1016/j.xpro.2025.104155
- Primary Topic
- Monoclonal and Polyclonal Antibodies Research
- Type
- article
- Field-Weighted Citation Impact
- 0.00