title: Methods for quantifying T cell receptor binding affinities and thermodynamics.
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.
authors: Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, Indiana, USA.
authors: Kurt H
authors: KH
authors: Piepenbrink
authors: Brian E
authors: BE
authors: Gloor
authors: Kathryn M
authors: KM
authors: Armstrong
authors: Brian M
authors: BM
authors: Baker
references: Nat Immunol. 2009 Feb;10(2):143-7
references: 19148199
references: Nat Biotechnol. 2005 Mar;23(3):349-54
references: 15723046
references: Methods Enzymol. 1995;259:194-221
references: 8538455
references: Nature. 1996 Nov 14;384(6605):134-41
references: 8906788
references: Methods Enzymol. 2003;361:505-29
references: 12624926
references: Methods Enzymol. 1996;274:492-503
references: 8902827
references: Immunity. 2008 Feb;28(2):171-82
references: 18275829
references: Biochemistry. 1996 Jul 9;35(27):8800-4
references: 8688415
references: Nat Immunol. 2003 Jul;4(7):657-63
references: 12796775
references: Biophys J. 1996 Oct;71(4):2049-55
references: 8889179
references: Curr Biol. 1993 Oct 1;3(10):658-67
references: 15335856
references: J Exp Med. 2005 Apr 18;201(8):1243-55
references: 15837811
references: J Immunol Methods. 2006 Jul 31;314(1-2):38-53
references: 16844138
references: Anal Biochem. 1989 May 15;179(1):131-7
references: 2757186
references: Mol Cell. 2003 Dec;12(6):1367-78
references: 14690592
references: Biophys J. 2006 Feb 15;90(4):1338-49
references: 16326918
references: Anal Biochem. 2008 Feb 15;373(2):395-7
references: 17920027
references: J Mol Recognit. 2008 Nov-Dec;21(6):355-400
references: 18951413
references: Biochemistry. 2002 Jun 11;41(23):7501-7
references: 12044184
references: J Immunol. 1996 Dec 15;157(12):5403-10
references: 8955188
references: J Biol Chem. 2005 Jan 21;280(3):1882-92
references: 15531581
references: Science. 1994 Feb 11;263(5148):777-84
references: 8303294
references: Science. 1994 Aug 12;265(5174):946-9
references: 8052850
references: J Mol Recognit. 1999 Sep-Oct;12(5):279-84
references: 10556875
references: J Mol Biol. 1995 Apr 28;248(2):478-86
references: 7739054
references: Proc Natl Acad Sci U S A. 1992 Apr 15;89(8):3429-33
references: 1565634
references: Methods Cell Biol. 2008;84:79-113
references: 17964929
references: Biophys J. 2002 Jun;82(6):3289-304
references: 12023252
references: J Mol Recognit. 2008 Jul-Aug;21(4):275-87
references: 18496839
references: J Am Chem Soc. 2003 Dec 3;125(48):14859-66
references: 14640663
references: EMBO J. 2007 Mar 21;26(6):1681-90
references: 17332746
references: Proc Natl Acad Sci U S A. 1994 Dec 20;91(26):12862-6
references: 7809136
references: Semin Immunol. 2007 Aug;19(4):216-24
references: 17398114
references: Immunol Res. 2008;41(3):267-94
references: 18726714
references: Proc Natl Acad Sci U S A. 2007 Oct 16;104(42):16639-44
references: 17921250
references: J Immunol Methods. 2001 Sep 1;255(1-2):57-66
references: 11470286
references: Biochemistry. 2008 Nov 25;47(47):12398-408
references: 18973345
references: Immunity. 1999 Mar;10(3):357-65
references: 10204491
references: Chem Rev. 1997 Aug 5;97(5):1233-1250
references: 11851449
references: Methods Enzymol. 1998;295:294-315
references: 9750224
references: Biophys J. 2007 Jul 15;93(2):597-609
references: 17449678
references: Methods Mol Biol. 2005;305:301-22
references: 15940004
references: Proteins. 1998 Nov 1;33(2):159-66
references: 9779785
references: Anal Biochem. 1998 Aug 1;261(2):203-10
references: 9716423
references: Biochemistry. 2001 Feb 13;40(6):1774-8
references: 11327839
references: Annu Rev Immunol. 1998;16:523-44
references: 9597140
references: Immunity. 1998 Apr;8(4):403-11
references: 9586631
references: Nat Biotechnol. 2000 Jul;18(7):754-9
references: 10888844
references: J Mol Biol. 2007 Oct 19;373(2):315-27
references: 17825839
references: Biochemistry. 2003 May 6;42(17):4954-61
references: 12718537
references: J Mol Biol. 1997 May 2;268(2):557-69
references: 9159490
references: J Mol Biol. 2005 Feb 18;346(2):533-50
references: 15670602
references: Immunity. 1999 Jul;11(1):45-56
references: 10435578
references: Annu Rev Immunol. 2006;24:419-66
references: 16551255
references: J Mol Biol. 2006 Oct 13;363(1):228-43
references: 16962135
references: J Biol Chem. 2004 Jul 9;279(28):29175-84
references: 15131131
references: Immunity. 2003 Oct;19(4):595-606
references: 14563323