Humanized antibodies
Protein display: K20

Amino acid sequences of the IGK V-J-REGION and IGH V-D-J-REGION of K20, an anti-b1 integrin chain monoclonal antibody [1,2] are displayed, according to the IMGT unique numbering for V-DOMAIN.
The humanized HuK20 has been expressed in cassette baculovirus vector for the production in insect cells [3,4].

Part 1- Humanization of the IGK V-J-REGION

______________________________________________________________________________________________________________________________________________________________________
IGK V-J-REGION                            FR1-IMGT           CDR1-IMGT       FR2-IMGT     CDR2-IMGT                 FR3-IMGT                   CDR3-IMGT    FR4-IMGT
                                           (1-26)             (27-38)        (39-55)       (56-65)                  (66-104)                   (105-117)    (118-128)
_______________________________ __________________________ ____________ _________________ __________ _______________________________________ _____________ ____________
                                1       10        20         30         40        50         60         70        80        90       100        110        120
                                .........|.........|...... ...|........ .|.........|..... ....|..... ....|.........|.........|.........|.... .....|....... ..|.........
Mouse MuVKK20            (1)    DIQLTQSPSSLSASLGGKVTITCKAS QDINKY...... IAWYQHEPGKGPRLLIR YTS....... KLESGIP.SRFSGSG..SGRDYSFSISNLEPEDIATYYC LQYY.....NLWT FGGGTKLEIKRK
Human REI                (2)    DIQLTQSPSSLSASVGDRVTITCQAS QDIIKY...... INWYQQKPGKAPKLLIY EAS....... NLQAGVP.SRFSGSG..SGTDFTFTISSLQPEDIATYYC QQYQS....LPYT FGQGTKVVIKRK
Humanized HuVKK20               DIQLTQSPSSLSASVGDRVTITCKAS QDINKY...... IAWYQQKPGKAPKLLIR YTS....... KLESGVP.SRFSGSG..SGTDFTFTISSLQPEDIATYYC LQYY.....NLWT FGQGTKVVIKRK

Legend: IMGT notes:
(1)The MuVKK20 sequence was run against the IMGT reference directory mouse Ig set, using IMGT/V-QUEST.
The closest IGK V-J rearrangement is the following: IGKV19-93*01-IGKJ1*01.
(2)The probable IGK V-J Rei rearrangement is the following: IGKV1-33*01 (or IGKV1D-33*01)-IGKJ2*01, as identified by IMGT curators (M. Ruiz, O. Posukh and M.-P. Lefranc Marie-Paule.Lefranc@igh.cnrs.fr).

Part 2- Humanization of the IGH V-D-J-REGION

______________________________________________________________________________________________________________________________________________________________________
IGH V-D-J-REGION                          FR1-IMGT           CDR1-IMGT       FR2-IMGT     CDR2-IMGT                 FR3-IMGT                   CDR3-IMGT    FR4-IMGT
                                           (1-26)             (27-38)        (39-55)       (56-65)                  (66-104)                   (105-117)    (118-128)
_______________________________ __________________________ ____________ _________________ __________ _______________________________________ _____________ ___________
                                1       10        20         30         40        50         60         70        80        90       100        110        120
                                .........|.........|...... ...|........ .|.........|..... ....|..... ....|.........|.........|.........|.... .....|....... ..|........
Mouse MuVHK20                   QVQLQESGT.ELVRPGASVKLSCKAS GYTFTDYY.... ISWVKQRPGQGLEWIAR IYPGSGNT.. FYNEKFK.GKATLTAETSSNTAYMQLSSLTSEDSAVYFC AIYYG....SGDY WGQGTTVTVSS
Human VHF1            (1)       QVQLQQSGT.EVKKPGASVKVSCKAS GYTFTLA..... MHWVRQAPGQGLEWMGW INGGNGET.. KSSQKFQ.GRVTITRDTSASTVYMELSSLRSEDTAIYYC ARDRWQPVVYFDT WGQGTTVTVSS
Humanized HuVHK20     (2)       QVQLQQSGT.EVKKPGASVKVSCKAS GYTFTDYY.... ISWVRQAPGQGLEWMGR IYPGSGNT.. FYNEKFK.GRVTITRDTSASTVYMELSSLRSEDTAIYYC AIYYG....SGDY WGQGTTVTVSS

IMGT notes:
(1)G54, R80 and D81 from MuVHK20 have to be conserved for HuVHK20 construction, because their substitution might enhance the immunogenicity of the humanized antibody.
(2)HuVHK20 is deleted of R106, in order to not affect the antigen binding site.

Reference:
[1]Poul, M.-A. et al., Molecular Immunology, 32, 101-106 (1995) PMID:7532786.
[2]Poul, M.-A. and Lefranc M.-P., Annals of the New York Academy of Sciences, 764, 359-361 (1995) PMID:7486546.
[3]Poul, M.-A. et al., Eur. J. Immunol. 25, 2005-2009 (1995) PMID:7542600.
[4]Poul, M.-A. et al., Immunotechnology, 1, 189-196 (1995) PMID:9373347.


Created: 07/06/2001
Author: Lisa Truong

Software material and data coming from IMGT server may be used for academic research only, provided that it is referred to IMGT®, and cited as "IMGT®, the international ImMunoGeneTics information system® http://www.imgt.org (founder and director: Marie-Paule Lefranc, Montpellier, France)." References to cite: Lefranc, M.-P. et al., Nucleic Acids Res., 27:209-212 (1999); doi: 10.1093/nar/27.1.209 Full text Cover; Ruiz, M. et al., Nucleic Acids Res., 28:219-221 (2000); doi: 10.1093/nar/28.1.219 Full text; Lefranc, M.-P., Nucleic Acids Res., 29:207-209 (2001); doi: 10.1093/nar/29.1.207 Full text; Lefranc, M.-P., Nucleic Acids Res., 31:307-310 (2003); doi: 10.1093/nar/gkg085 Full text; Lefranc, M.-P. et al., In Silico Biol., 5, 0006 (2004) [Epub], 5:45-60 (2005); Lefranc, M.-P. et al., Nucleic Acids Res., 33:D593-597 (2005); doi: 10.1093/nar/gki065 Full text; Lefranc, M.-P. et al., Nucleic Acids Res., 37:D1006-1012 (2009); doi: 10.1093/nar/gkn838 Full text; Lefranc, M.-P. et al., Nucleic Acids Res., 43:D413-422 (2015); doi: 10.1093/nar/gku1056 Full text.
For any other use please contact Marie-Paule Lefranc Marie-Paule.Lefranc@igh.cnrs.fr.

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