The immunogenicity of humanized and fully human antibodies: residual immunogenicity resides in the CDR regions.
Monoclonal antibodies represent an attractive therapeutic tool as they are highly specific for their targets, convey effector functions and enjoy robust manufacturing procedures. Humanization of murine monoclonal antibodies has vastly improved their in vivo tolerability. Humanization, the replacement of mouse constant regions and V framework regions for human sequences, results in a significantly less immunogenic product. However, some humanized and even fully human sequence-derived antibody molecules still carry immunological risk. To more fully understand the immunologic potential of humanized and human antibodies, we analyzed CD4(+) helper T cell epitopes in a set of eight humanized antibodies. The antibodies studied represented a number of different VH and VL family members carrying unique CDR regions. In spite of these differences, CD4(+) T cell epitopes were found only in CDR-sequence containing regions. We were able to incorporate up to two amino acid modifications in a single epitope that reduced the immunogenic potential while retaining full biologic function. We propose that immunogenicity will always be present in some antibody molecules due to the nature of the antigen-specific combining sites. A consequence of this result is modifications to reduce immunogenicity will be centered on the affinity-determining regions. Modifications to CDR regions can be designed that reduce the immunogenic potential while maintaining the bioactivity of the antibody molecule.
Harding
Fiona A
FA
Facet Biotech, Redwood City, CA, USA. fiona.harding@facetbiotech.com
Stickler
Marcia M
MM
Razo
Jennifer
J
DuBridge
Robert B
RB
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology*; D000915:Antibody Affinity; D000918:Antibody Specificity; D002478:Cells, Cultured; D022801:Complementarity Determining Regions / Q000235:genetics / Q000276:immunology*; D018984:Epitopes, T-Lymphocyte / Q000235:genetics / Q000276:immunology; D006801:Humans; D051379:Mice; D015202:Protein Engineering; D006377:T-Lymphocytes, Helper-Inducer / Q000276:immunology
D000911:Antibodies, Monoclonal; D022801:Complementarity Determining Regions; D018984:Epitopes, T-Lymphocyte
Nat Rev Immunol. 2005 Nov;5(11):853-65
16261173
Methods. 2005 May;36(1):3-10
15848070
J Clin Invest. 2004 May;113(10):1498-510
15146248
Immunol Cell Biol. 2008 Feb;86(2):146-52
18227854
J Clin Invest. 1998 Jul 1;102(1):215-22
9649575
Transplantation. 2006 Feb 15;81(3):308-15
16477212
Toxicology. 1995 Dec 20;105(1):23-9
8638282
Semin Hematol. 2004 Jan;41(1 Suppl 1):82-8
14872427
J Exp Med. 2004 Oct 4;200(7):847-56
15466619
Nat Rev Immunol. 2008 Jul;8(7):523-32
18566595
Aliment Pharmacol Ther. 2008 Nov 1;28(9):1122-6
18691349
Curr Opin Immunol. 1998 Apr;10(2):208-13
9602310
Biotechnol Annu Rev. 2008;14:191-202
18606364
Curr Opin Pharmacol. 2008 Oct;8(5):620-6
18775515
J Exp Med. 2009 Aug 3;206(8):1643-6
19635860
J Immunol. 2004 Jun 15;172(12):7476-84
15187126
J Immunol. 1990 Jun 1;144(11):4256-61
2341720
Cell. 1998 Dec 23;95(7):875-8
9875841
Proc Natl Acad Sci U S A. 2004 Jun 22;101(25):9205-10
15197260
Genes Immun. 2004 Jan;5(1):1-7
14735143
J Immunol. 1987 Oct 15;139(8):2825-33
3498771
Nat Biotechnol. 2005 Sep;23(9):1105-16
16151404
J Immunol. 2005 Aug 15;175(4):2391-400
16081810
Rheumatol Int. 2007 Jan;27(3):269-74
17006705
Curr Opin Immunol. 2009 Jun;21(3):266-73
19502021
Proc Natl Acad Sci U S A. 2008 Apr 8;105(14):5471-6
18367670
Ann Oncol. 2008 Jan;19(1):92-8
17785764
J Immunol. 1996 Aug 1;157(3):1037-46
8757607
Scand J Immunol. 2009 Dec;70(6):526-30
19906193
Ann Rheum Dis. 2009 Nov;68(11):1739-45
19019895
Transplantation. 1990 Apr;49(4):697-702
2109379
Drug Discov Today. 2004 Jan 15;9(2):82-90
15012932
Arthritis Rheum. 2008 Nov;58(11):3402-12
18975305
Methods. 2005 May;36(1):69-83
15848076
Dev Biol (Basel). 2003;112:15-21
12762500
Clin Exp Immunol. 2002 Jul;129(1):169-76
12100037
Anal Biochem. 1997 Apr 5;247(1):102-10
9126378
Annu Rev Immunol. 2006;24:541-70
16551259
N Engl J Med. 2008 Mar 13;358(11):1109-17
18337601
J Immunother. 2000 Nov-Dec;23(6):654-60
11186153
Curr Med Chem. 2002 Dec;9(24):2191-9
12470241
Immunol Rev. 2004 Aug;200:182-96
15242405
Trends Immunol. 2002 Nov;23(11):526-9
12401404
Curr Opin Drug Discov Devel. 2007 Mar;10(2):219-27
17436557
AAPS J. 2006 Aug 04;8(3):E501-7
17025268
Thymus. 1994;22(3):141-52
7940642
Rheumatology (Oxford). 2008 May;47(5):567-71
18316337
Annu Rev Immunol. 2007;25:21-50
17029568
Nat Biotechnol. 2007 Oct;25(10):1134-43
17921999
Blood. 2008 Mar 1;111(5):2725-32
18156498
Curr Opin Immunol. 2008 Aug;20(4):450-9
18606226
J Neuroimmunol. 2009 Jul 25;212(1-2):65-73
19477024
Adv Immunol. 2007;95:83-110
17869611
J Immunol Methods. 1999 Dec 10;231(1-2):11-23
10648924
Curr Opin Mol Ther. 2003 Apr;5(2):172-9
12772508
Jpn J Clin Oncol. 2009 May;39(5):321-6
19287023
Nat Immunol. 2000 Dec;1(6):453-5
11101860
Blood Rev. 2009 May;23(3):137-42
18951668
Dev Biol (Basel). 2003;112:55-9
12762504
Nat Biotechnol. 2005 Sep;23(9):1117-25
16151405
Clin Exp Immunol. 2004 Sep;137(3):496-502
15320898
J Immunol Methods. 2003 Oct 1;281(1-2):95-108
14580884
Clin Cancer Res. 2003 Apr;9(4):1347-53
12684403
J Immunol. 2005 Aug 15;175(4):2184-90
16081785
Immunity. 2009 May;30(5):646-55
19464987
Cancer Res. 2001 Sep 15;61(18):6851-9
11559561
Transfusion. 2006 Sep;46(9):1526-36
16965580
Vaccine. 2009 Dec 10;28(1):14-20
19818425
J Immunol. 2007 Jun 1;178(11):7467-72
17513798
Immunotechnology. 1997 Mar;3(1):31-43
9154466
Annu Rev Immunol. 2001;19:23-45
11244029
Ann N Y Acad Sci. 1995 Sep 29;764:242-60
7486531
Nature. 1994 Apr 28;368(6474):856-9
8159246
Ann Rheum Dis. 2006 Sep;65(9):1249-50
16905585
Int Immunol. 2007 Jul;19(7):857-65
17576752
Semin Thromb Hemost. 2000;26(2):151-5
10919407
Mol Cancer Ther. 2005 Nov;4(11):1791-800
16276001
Nat Rev Immunol. 2009 Dec;9(12):833-44
19935803
N Engl J Med. 1992 Oct 22;327(17):1209-15
1406793
J Mol Recognit. 2003 Sep-Oct;16(5):299-317
14523943
Autoimmunity. 2007 Mar;40(2):131-7
17453712
Neurol Sci. 2002 Apr;23 Suppl 1:S1-8
12032582
Clin Exp Rheumatol. 1999 Nov-Dec;17(6 Suppl 18):S73-7
10589362
Haematologica. 2000 Oct;85(10 Suppl):2-5; discussion 5-6
11187864
IDrugs. 2009 Apr;12(4):233-7
19350467
Ann Immunol (Paris). 1974 Jan;125C(1-2):373-89
4142565
Curr Opin Biotechnol. 2009 Dec;20(6):708-14
19880308
Scand J Immunol. 1999 Sep;50(3):296-301
10447939
Arthritis Rheum. 2008 Apr;58(4):964-75
18383539
Dev Biol (Basel). 2003;112:37-53
12762503
Clin Exp Immunol. 2008 Apr;152(1):138-46
18307520