Methods for quantifying T cell receptor binding affinities and thermodynamics.

Kurt H. Piepenbrink, Brian E. Gloor, Kathryn M. Armstrong, Brian M. Baker

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

αβ T cell receptors (TCRs) recognize peptide antigens bound and presented by class I or class II major histocompatibility complex (MHC) proteins. Recognition of a peptide/MHC complex is required for initiation and propagation of a cellular immune response, as well as the development and maintenance of the T cell repertoire. Here, we discuss methods to quantify the affinities and thermodynamics of interactions between soluble ectodomains of TCRs and their peptide/MHC ligands, focusing on titration calorimetry, surface plasmon resonance, and fluorescence anisotropy. As TCRs typically bind ligand with weak-to-moderate affinities, we focus the discussion on means to enhance the accuracy and precision of low-affinity measurements. In addition to further elucidating the biology of the T cell mediated immune response, more reliable low-affinity measurements will aid with more probing studies with mutants or altered peptides that can help illuminate the physical underpinnings of how TCRs achieve their remarkable recognition properties.

Original languageEnglish (US)
Pages (from-to)359-381
Number of pages23
JournalMethods in enzymology
Volume466
StatePublished - Dec 1 2009

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T-Cell Antigen Receptor
Thermodynamics
Major Histocompatibility Complex
Peptides
T-cells
Ligands
T-Lymphocytes
Calorimetry
Fluorescence Polarization
Surface Plasmon Resonance
Surface plasmon resonance
Titration
Cellular Immunity
Anisotropy
Fluorescence
Maintenance
Antigens
Proteins

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Methods for quantifying T cell receptor binding affinities and thermodynamics. / Piepenbrink, Kurt H.; Gloor, Brian E.; Armstrong, Kathryn M.; Baker, Brian M.

In: Methods in enzymology, Vol. 466, 01.12.2009, p. 359-381.

Research output: Contribution to journalArticle

Piepenbrink, Kurt H. ; Gloor, Brian E. ; Armstrong, Kathryn M. ; Baker, Brian M. / Methods for quantifying T cell receptor binding affinities and thermodynamics. In: Methods in enzymology. 2009 ; Vol. 466. pp. 359-381.
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