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

Matias Cardenas; Sasha Compton; Ashkon Hunt; Pradeep Chopra; Daniel R Perez; Geert-Jan Boons; Daniela S Rajao

doi:10.1016/j.xpro.2025.104155

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

Matias Cardenas^*
, Sasha Compton
, Ashkon Hunt
, Pradeep Chopra
, Daniel R Perez
, Geert-Jan Boons
, Daniela S Rajao^*

^*Corresponding author for this work

University of Georgia

Research output: Contribution to journal › Article › Academic › peer-review

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.

Original language	English
Article number	104155
Number of pages	24
Journal	STAR Protocols
Volume	6
Issue number	4
Early online date	23 Oct 2025
DOIs	https://doi.org/10.1016/j.xpro.2025.104155
Publication status	Published - 19 Dec 2025

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10.1016/j.xpro.2025.104155Licence: CC BY-NC-ND

Protocol to measure the kinetics of MHC class II-influenza A binding using biolayer interferometryFinal published version, 8.48 MBLicence: CC BY-NC-ND

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

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.",

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AU - Cardenas, Matias

AU - Compton, Sasha

AU - Hunt, Ashkon

AU - Chopra, Pradeep

AU - Perez, Daniel R

AU - Boons, Geert-Jan

AU - Rajao, Daniela S

PY - 2025/12/19

Y1 - 2025/12/19

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AB - 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.

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DO - 10.1016/j.xpro.2025.104155

M3 - Article

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JO - STAR Protocols

JF - STAR Protocols

IS - 4

M1 - 104155

ER -

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

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