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

Area of Science:

• Immunology
• Virology
• Biochemistry

Background:

• Major histocompatibility complex (MHC) class II acts as a functional receptor for certain influenza A virus (FLUAV) subtypes.
• Understanding the binding kinetics between MHC class II and FLUAV is crucial for virological research.

Purpose of the Study:

• To present a novel protocol for quantifying the binding kinetics of MHC class II and influenza A virus interactions.
• To enable the screening of diverse FLUAV subtypes for their binding affinity to MHC class II.

Main Methods:

• Production of recombinant human leukocyte antigen DR isotype (HLA-DR) MHC class II utilizing a baculovirus expression system.
• Modification of HLA-DR for use as a biolayer interferometry (BLI) probe.
• Kinetic analysis of whole FLUAV viruses binding to the prepared BLI probe.

Main Results:

• A detailed protocol for measuring MHC class II-influenza A binding kinetics was successfully established.
• The method allows for the characterization of binding affinities between different FLUAV subtypes and MHC class II.

Conclusions:

• The developed biolayer interferometry protocol provides a robust method for studying MHC class II-influenza A virus interactions.
• This technique facilitates the screening and characterization of various influenza A virus subtypes' affinity for MHC class II, advancing our understanding of host-pathogen interactions.