title: Removal of B cell epitopes as a practical approach for reducing the immunogenicity of foreign protein-based therapeutics.
abstract: Immunogenicity of non-human proteins with useful therapeutic properties has prevented their development for use in the therapy of disease. However, this class of proteins could be very useful, if their immunogenicity could be markedly reduced so that many treatment cycles could be administered. One approach to reduce the immunogenicity of foreign proteins is to identify B cell epitopes on the protein and eliminate them by mutagenesis. In this article, theoretical aspects and experimental evidence for the feasibility of B cell epitope removal is reviewed. A special focus is given to our results with deimmunization of recombinant immunotoxins in which Fvs are fused to a 38kDa portion of the bacterial protein, Pseudomonas exotoxin A (PE38). Immunotoxins targeting CD22 and CD25 have produced complete remissions in many patients with drug resistant Hairy Cell Leukemia and are being evaluated in other malignancies. Experimental data summarized in this review indicates that removal of B cell epitopes is a practical approach for making less immunogenic protein therapeutics from non-human functional proteins. This approach requires grouping of the epitopes to identify targets for deimmunization followed by quantitative analysis of the decrease in affinity produced by the mutations in B cell epitopes.
authors.affiliation: Cancer Biology Research Center, Sanford Research/USD, Sioux Falls, SD 57105, USA. nagatas@sanfordhealth.org
authors.forename: Satoshi
authors.initials: S
authors.lastname: Nagata
authors.forename: Ira
authors.initials: I
authors.lastname: Pastan
mesh_terms: D000818:Animals; D000915:Antibody Affinity; D000937:Antigen-Antibody Reactions; D018985:Epitopes, B-Lymphocyte; D018984:Epitopes, T-Lymphocyte; D006801:Humans; D000922:Immunotoxins / Q000276:immunology; D016296:Mutagenesis; D017354:Point Mutation; D011487:Protein Conformation; D011506:Proteins / Q000276:immunology*
chemical_list: D018985:Epitopes, B-Lymphocyte; D018984:Epitopes, T-Lymphocyte; D000922:Immunotoxins; D011506:Proteins
references.citation: Nat Rev Immunol. 2007 Jun;7(6):485-90
references.pmid: 17479127
references.citation: Immunol Cell Biol. 2001 Oct;79(5):507-11
references.pmid: 11564159
references.citation: Curr Opin Mol Ther. 2004 Feb;6(1):10-6
references.pmid: 15011776
references.citation: Adv Immunol. 2000;75:159-208
references.pmid: 10879284
references.citation: J Immunol. 1975 Apr;114(4):1302-6
references.pmid: 46896
references.citation: Annu Rev Immunol. 1993;11:331-60
references.pmid: 8476565
references.citation: Nature. 1986 May 29-Jun 4;321(6069):522-5
references.pmid: 3713831
references.citation: Biochemistry. 1964 Jul;3:996-1008
references.pmid: 14214095
references.citation: J Mol Biol. 1971 Feb 14;55(3):379-400
references.pmid: 5551392
references.citation: J Immunol Methods. 1992 Feb 14;147(1):125-34
references.pmid: 1311735
references.citation: Methods Mol Biol. 1996;66:55-66
references.pmid: 8959704
references.citation: Mol Immunol. 2007 Feb;44(5):1029-41
references.pmid: 16647121
references.citation: Immunol Invest. 1992 Oct;21(6):523-38
references.pmid: 1385323
references.citation: Immunol Invest. 1997 Jun;26(4):473-89
references.pmid: 9246567
references.citation: Proc Natl Acad Sci U S A. 2005 May 31;102(22):7946-51
references.pmid: 15905329
references.citation: Clin Exp Immunol. 1972 Sep;12(1):121-32
references.pmid: 5083460
references.citation: Am J Obstet Gynecol. 2007 Jan;196(1):7-15
references.pmid: 17240220
references.citation: Science. 1987 Mar 6;235(4793):1184-90
references.pmid: 3823878
references.citation: Immunogenetics. 2005 Jun;57(5):326-36
references.pmid: 15895191
references.citation: J Mol Recognit. 2002 Sep-Oct;15(5):240-7
references.pmid: 12447900
references.citation: Cell. 1990 May 18;61(4):553-6
references.pmid: 1693095
references.citation: J Immunol Methods. 1997 Aug 7;206(1-2):43-52
references.pmid: 9328567
references.citation: Assay Drug Dev Technol. 2007 Oct;5(5):655-62
references.pmid: 17939757
references.citation: Proc Natl Acad Sci U S A. 2000 Mar 14;97(6):2719-24
references.pmid: 10716997
references.citation: Nucleic Acids Res. 2006 Jan 1;34(Database issue):D777-80
references.pmid: 16381978
references.citation: N Engl J Med. 2001 Jul 26;345(4):241-7
references.pmid: 11474661
references.citation: J Immunol. 1987 Jan 1;138(1):213-9
references.pmid: 2431055
references.citation: Semin Immunol. 1989 Sep;1(1):5-12
references.pmid: 15630954
references.citation: Protein Sci. 2001 Mar;10(3):491-503
references.pmid: 11344318
references.citation: Methods Enzymol. 1991;203:176-201
references.pmid: 1722270
references.citation: Circulation. 1996 Jul 15;94(2):197-206
references.pmid: 8674179
references.citation: Expert Opin Biol Ther. 2006 Jan;6(1):1-7
references.pmid: 16370909
references.citation: Science. 1985 Sep 6;229(4717):932-40
references.pmid: 2410982
references.citation: Int J Cardiol. 2003 Oct;91(2-3):115-27
references.pmid: 14559121
references.citation: J Pharm Biomed Anal. 2008 Dec 15;48(5):1267-81
references.pmid: 18993008
references.citation: Int J Med Microbiol. 2004 Apr;293(7-8):577-82
references.pmid: 15149034
references.citation: J Immunol Methods. 1992 Jun 24;150(1-2):77-90
references.pmid: 1613260
references.citation: J Immunol. 2006 Dec 15;177(12):8822-34
references.pmid: 17142785
references.citation: Protein J. 2004 Jan;23(1):39-52
references.pmid: 15115181
references.citation: Anal Chem. 2001 Feb 1;73(3):471-80
references.pmid: 11217749
references.citation: J Theor Biol. 2004 Dec 21;231(4):535-48
references.pmid: 15488530
references.citation: Clin Ther. 2007 Apr;29(4):683-90
references.pmid: 17617291
references.citation: Nat Rev Drug Discov. 2008 Jan;7(1):21-39
references.pmid: 18097458
references.citation: Nature. 1986 Aug 21-27;322(6081):747-8
references.pmid: 2427953
references.citation: Thromb Haemost. 2008 May;99(5):874-82
references.pmid: 18449416
references.citation: Protein Sci. 1994 Dec;3(12):2351-7
references.pmid: 7538848
references.citation: Thromb Haemost. 1994 Aug;72(2):297-301
references.pmid: 7831668
references.citation: Proc Natl Acad Sci U S A. 2008 Aug 12;105(32):11311-6
references.pmid: 18678888
references.citation: J Immunotoxicol. 2006 Sep 1;3(3):151-6
references.pmid: 18958695
references.citation: J Mol Biol. 1992 Aug 5;226(3):851-65
references.pmid: 1380563
references.citation: Nature. 1984 Sep 13-19;311(5982):123-6
references.pmid: 6206398
references.citation: Anal Chem. 1997 Jan 15;69(2):175-82
references.pmid: 8997894
references.citation: J Mol Recognit. 2004 Mar-Apr;17(2):132-43
references.pmid: 15027033
references.citation: Protein Sci. 2005 Jan;14(1):246-8
references.pmid: 15576553
references.citation: J Exp Med. 1994 Nov 1;180(5):1889-900
references.pmid: 7964465
references.citation: Proc Natl Acad Sci U S A. 1996 Mar 19;93(6):2264-71
references.pmid: 8637861
references.citation: Amino Acids. 2007 Sep;33(3):423-8
references.pmid: 17252308
references.citation: Methods Mol Biol. 2009;524:3-20
references.pmid: 19377933
references.citation: Blood. 2000 Aug 15;96(4):1425-32
references.pmid: 10942387
references.citation: Annu Rev Biochem. 1990;59:439-73
references.pmid: 2197980
references.citation: Mol Immunol. 1997 Aug-Sep;34(12-13):843-53
references.pmid: 9464520
references.citation: Annu Rev Med. 2007;58:221-37
references.pmid: 17059365
references.citation: Nat Rev Cancer. 2006 Jul;6(7):559-65
references.pmid: 16794638
references.citation: J Immunol. 1992 Nov 15;149(10):3260-72
references.pmid: 1431104
references.citation: Clin Ther. 2002 Nov;24(11):1720-40; discussion 1719
references.pmid: 12501870
references.citation: J Immunol Methods. 2004 Sep;292(1-2):141-55
references.pmid: 15350519
references.citation: Transplantation. 2005 Feb 15;79(3 Suppl):S8-S11
references.pmid: 15699753
references.citation: Curr Protoc Immunol. 2004 May;Chapter 9:Unit 9.4
references.pmid: 18432936
references.citation: Adv Exp Med Biol. 2008;640:198-207
references.pmid: 19065793
references.citation: Nature. 1984 Dec 13-19;312(5995):643-6
references.pmid: 6095115
references.citation: Science. 1989 Jun 2;244(4908):1081-5
references.pmid: 2471267
references.citation: Nat Biotechnol. 2007 May;25(5):555-61
references.pmid: 17483842
references.citation: Orthop Rev. 1992 Sep;21(9):1034-42, 1130-1
references.pmid: 1437254
references.citation: Proc Natl Acad Sci U S A. 1984 Nov;81(21):6851-5
references.pmid: 6436822
references.citation: Biologicals. 2001 Sep-Dec;29(3-4):209-13
references.pmid: 11851317
references.citation: Methods Mol Biol. 2009;524:335-44
references.pmid: 19377956
references.citation: J Immunol Methods. 2003 Jul;278(1-2):127-44
references.pmid: 12957402
references.citation: Prog Clin Biol Res. 1980;47:107-17
references.pmid: 6163155
references.citation: J Clin Oncol. 2005 Sep 20;23(27):6719-29
references.pmid: 16061911
references.citation: Trends Biotechnol. 2006 Jun;24(6):274-80
references.pmid: 16631266
references.citation: Transplantation. 2006 Feb 15;81(3):308-15
references.pmid: 16477212
references.citation: Mol Immunol. 2009 Feb;46(5):840-7
references.pmid: 18947876
references.citation: Br J Cancer. 2004 Jun 14;90(12):2402-10
references.pmid: 15162148
references.citation: Scand J Immunol. 1996 Nov;44(5):478-84
references.pmid: 8947599
references.citation: Immunity. 1994 Jun;1(3):167-78
references.pmid: 7534202
references.citation: Annu Rev Immunol. 1994;12:117-39
references.pmid: 8011279
references.citation: J Mol Recognit. 2007 Mar-Apr;20(2):75-82
references.pmid: 17205610
references.citation: Protein Sci. 2006 Nov;15(11):2558-67
references.pmid: 17001032
references.citation: Immunol Invest. 1999 Mar-May;28(2-3):89-101
references.pmid: 10484684
references.citation: Blood. 2008 Jan 1;111(1):338-43
references.pmid: 17895404
references.citation: J Virol. 2005 Oct;79(19):12148-63
references.pmid: 16160142
references.citation: Int J Pharm. 2003 Nov 6;266(1-2):3-16
references.pmid: 14559389
references.citation: BMC Bioinformatics. 2008 Dec 02;9:514
references.pmid: 19055730