﻿               SEQUENCE LISTING

<110> Fund. Inst. de Investigación contra la Leucemia Josep
      Carreras (IJC) et al.

<120> CD22 targeting-moiety for the treatment of B-cell acute lymphoblastic leukemia (B-ALL).

<130> 906 025

<150> EP 20382175.6
<151> 2020-03-11

<160> 23

<170> BiSSAP 1.3.6

<210> 1
<211> 11
<212> PRT
<213> Artificial Sequence


<220> 
<221> PEPTIDE
<222> 1..11

<220> 
<223> LCDR1

<400> 1
Gln Ser Leu Leu Asp Ser Asp Gly Lys Thr Tyr 
1               5                   10      

<210> 2
<400> 2
000

<210> 3
<211> 9
<212> PRT
<213> Artificial Sequence


<220> 
<223> LCDR3

<400> 3
Trp Gln Gly Thr His Phe Pro Trp Thr 
1               5                   

<210> 4
<211> 8
<212> PRT
<213> Artificial Sequence


<220> 
<221> PEPTIDE
<222> 1..8

<220> 
<223> HCDR1

<400> 4
Gly Asp Ser Ile Thr Ser Gly Tyr 
1               5               

<210> 5
<211> 7
<212> PRT
<213> Artificial Sequence


<220> 
<221> PEPTIDE
<222> 1..7

<220> 
<223> HCDR2

<400> 5
Ile Ser Tyr Ser Gly Ser Thr 
1               5           

<210> 6
<211> 11
<212> PRT
<213> Artificial Sequence


<220> 
<221> PEPTIDE
<222> 1..11

<220> 
<223> HCDR3

<400> 6
Ala Arg Tyr Pro Ser Pro Asp Ala Met Asn Tyr 
1               5                   10      

<210> 7
<211> 114
<212> PRT
<213> Artificial Sequence


<220> 
<221> PEPTIDE
<222> 1..114

<220> 
<223> VL domain

<400> 7
Asp Val Val Met Thr Gln Thr Pro Leu Thr Leu Ser Val Thr Ile Gly 
1               5                   10                  15      
Gln Pro Ala Ser Ile Ser Cys Lys Ser Ser Gln Ser Leu Leu Asp Ser 
            20                  25                  30          
Asp Gly Lys Thr Tyr Leu Asn Trp Leu Leu Gln Arg Pro Gly Gln Ser 
        35                  40                  45              
Pro Lys Arg Leu Ile Tyr Leu Val Ser Lys Leu Asp Ser Gly Val Pro 
    50                  55                  60                  
Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 
65                  70                  75                  80  
Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys Trp Gln Gly 
                85                  90                  95      
Thr His Phe Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 
            100                 105                 110         
Arg Ala 
        

<210> 8
<211> 117
<212> PRT
<213> Artificial Sequence


<220> 
<221> PEPTIDE
<222> 1..117

<220> 
<223> VH domain

<400> 8
Glu Val Gln Leu Gln Glu Ser Gly Pro Ser Leu Val Lys Pro Ser Gln 
1               5                   10                  15      
Thr Leu Ser Leu Thr Cys Ser Val Thr Gly Asp Ser Ile Thr Ser Gly 
            20                  25                  30          
Tyr Trp Asn Trp Ile Arg Lys Phe Pro Gly Asn Lys Leu Glu Tyr Met 
        35                  40                  45              
Gly Tyr Ile Ser Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser Leu Lys 
    50                  55                  60                  
Ser Arg Ile Ser Ile Thr Arg Asp Thr Ser Lys Asn Gln Tyr Tyr Met 
65                  70                  75                  80  
Gln Leu Lys Ser Val Thr Thr Glu Asp Thr Ala Thr Tyr Tyr Cys Ala 
                85                  90                  95      
Arg Tyr Pro Ser Pro Asp Ala Met Asn Tyr Trp Gly Gln Gly Thr Ser 
            100                 105                 110         
Val Thr Val Ser Ser 
        115         

<210> 9
<211> 251
<212> PRT
<213> Artificial Sequence


<220> 
<221> PEPTIDE
<222> 1..251

<220> 
<223> scFv derived from clone hCD22.7

<400> 9
Glu Val Gln Leu Gln Glu Ser Gly Pro Ser Leu Val Lys Pro Ser Gln 
1               5                   10                  15      
Thr Leu Ser Leu Thr Cys Ser Val Thr Gly Asp Ser Ile Thr Ser Gly 
            20                  25                  30          
Tyr Trp Asn Trp Ile Arg Lys Phe Pro Gly Asn Lys Leu Glu Tyr Met 
        35                  40                  45              
Gly Tyr Ile Ser Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser Leu Lys 
    50                  55                  60                  
Ser Arg Ile Ser Ile Thr Arg Asp Thr Ser Lys Asn Gln Tyr Tyr Met 
65                  70                  75                  80  
Gln Leu Lys Ser Val Thr Thr Glu Asp Thr Ala Thr Tyr Tyr Cys Ala 
                85                  90                  95      
Arg Tyr Pro Ser Pro Asp Ala Met Asn Tyr Trp Gly Gln Gly Thr Ser 
            100                 105                 110         
Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 
        115                 120                 125             
Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Val Val Met Thr Gln Thr 
    130                 135                 140                 
Pro Leu Thr Leu Ser Val Thr Ile Gly Gln Pro Ala Ser Ile Ser Cys 
145                 150                 155                 160 
Lys Ser Ser Gln Ser Leu Leu Asp Ser Asp Gly Lys Thr Tyr Leu Asn 
                165                 170                 175     
Trp Leu Leu Gln Arg Pro Gly Gln Ser Pro Lys Arg Leu Ile Tyr Leu 
            180                 185                 190         
Val Ser Lys Leu Asp Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Gly 
        195                 200                 205             
Ser Gly Thr Asp Phe Thr Leu Lys Ile Ser Arg Val Glu Ala Glu Asp 
    210                 215                 220                 
Leu Gly Val Tyr Tyr Cys Trp Gln Gly Thr His Phe Pro Trp Thr Phe 
225                 230                 235                 240 
Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Ala 
                245                 250     

<210> 10
<211> 69
<212> PRT
<213> Artificial Sequence


<220> 
<221> PEPTIDE
<222> 1..69

<220> 
<223> Transmembrane domain derived from CD8

<400> 10
Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala 
1               5                   10                  15      
Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly 
            20                  25                  30          
Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile 
        35                  40                  45              
Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val 
    50                  55                  60                  
Ile Thr Leu Tyr Cys 
65                  

<210> 11
<211> 113
<212> PRT
<213> Artificial Sequence


<220> 
<221> PEPTIDE
<222> 1..113

<220> 
<223> Intracellular signaling domain derived from CD3

<400> 11
Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly 
1               5                   10                  15      
Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr 
            20                  25                  30          
Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys 
        35                  40                  45              
Pro Gln Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln 
    50                  55                  60                  
Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu 
65                  70                  75                  80  
Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr 
                85                  90                  95      
Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro 
            100                 105                 110         
Arg 
    

<210> 12
<211> 42
<212> PRT
<213> Artificial Sequence


<220> 
<221> PEPTIDE
<222> 1..42

<220> 
<223> Costimulatory signaling domain derived from CD137

<400> 12
Lys Arg Gly Arg Lys Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met 
1               5                   10                  15      
Arg Pro Val Gln Thr Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe 
            20                  25                  30          
Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu 
        35                  40          

<210> 13
<211> 475
<212> PRT
<213> Artificial Sequence


<220> 
<221> PEPTIDE
<222> 1..475

<220> 
<223> Full sequence of the CAR

<400> 13
Glu Val Gln Leu Gln Glu Ser Gly Pro Ser Leu Val Lys Pro Ser Gln 
1               5                   10                  15      
Thr Leu Ser Leu Thr Cys Ser Val Thr Gly Asp Ser Ile Thr Ser Gly 
            20                  25                  30          
Tyr Trp Asn Trp Ile Arg Lys Phe Pro Gly Asn Lys Leu Glu Tyr Met 
        35                  40                  45              
Gly Tyr Ile Ser Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser Leu Lys 
    50                  55                  60                  
Ser Arg Ile Ser Ile Thr Arg Asp Thr Ser Lys Asn Gln Tyr Tyr Met 
65                  70                  75                  80  
Gln Leu Lys Ser Val Thr Thr Glu Asp Thr Ala Thr Tyr Tyr Cys Ala 
                85                  90                  95      
Arg Tyr Pro Ser Pro Asp Ala Met Asn Tyr Trp Gly Gln Gly Thr Ser 
            100                 105                 110         
Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 
        115                 120                 125             
Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Val Val Met Thr Gln Thr 
    130                 135                 140                 
Pro Leu Thr Leu Ser Val Thr Ile Gly Gln Pro Ala Ser Ile Ser Cys 
145                 150                 155                 160 
Lys Ser Ser Gln Ser Leu Leu Asp Ser Asp Gly Lys Thr Tyr Leu Asn 
                165                 170                 175     
Trp Leu Leu Gln Arg Pro Gly Gln Ser Pro Lys Arg Leu Ile Tyr Leu 
            180                 185                 190         
Val Ser Lys Leu Asp Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Gly 
        195                 200                 205             
Ser Gly Thr Asp Phe Thr Leu Lys Ile Ser Arg Val Glu Ala Glu Asp 
    210                 215                 220                 
Leu Gly Val Tyr Tyr Cys Trp Gln Gly Thr His Phe Pro Trp Thr Phe 
225                 230                 235                 240 
Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Ala Thr Thr Thr Pro Ala 
                245                 250                 255     
Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser 
            260                 265                 270         
Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr 
        275                 280                 285             
Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala 
    290                 295                 300                 
Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys 
305                 310                 315                 320 
Lys Arg Gly Arg Lys Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met 
                325                 330                 335     
Arg Pro Val Gln Thr Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe 
            340                 345                 350         
Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu Arg Val Lys Phe Ser Arg 
        355                 360                 365             
Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn 
    370                 375                 380                 
Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg 
385                 390                 395                 400 
Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Gln Arg Arg Lys Asn 
                405                 410                 415     
Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu 
            420                 425                 430         
Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly 
        435                 440                 445             
His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr 
    450                 455                 460                 
Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg 
465                 470                 475 

<210> 14
<211> 119
<212> PRT
<213> Artificial Sequence


<220> 
<221> PEPTIDE
<222> 1..119

<220> 
<223> The first Ig extracellular domain of the CD22 antigen, the
      farthest domain from the membrane

<400> 14
Asp Ser Ser Lys Trp Val Phe Glu His Pro Glu Thr Leu Tyr Ala Trp 
1               5                   10                  15      
Glu Gly Ala Cys Val Trp Ile Pro Cys Thr Tyr Arg Ala Leu Asp Gly 
            20                  25                  30          
Asp Leu Glu Ser Phe Ile Leu Phe His Asn Pro Glu Tyr Asn Lys Asn 
        35                  40                  45              
Thr Ser Lys Phe Asp Gly Thr Arg Leu Tyr Glu Ser Thr Lys Asp Gly 
    50                  55                  60                  
Lys Val Pro Ser Glu Gln Lys Arg Val Gln Phe Leu Gly Asp Lys Asn 
65                  70                  75                  80  
Lys Asn Cys Thr Leu Ser Ile His Pro Val His Leu Asn Asp Ser Gly 
                85                  90                  95      
Gln Leu Gly Leu Arg Met Glu Ser Lys Thr Glu Lys Trp Met Glu Arg 
            100                 105                 110         
Ile His Leu Asn Val Ser Glu 
        115                 

<210> 15
<211> 9
<212> PRT
<213> Artificial Sequence


<220> 
<221> PEPTIDE
<222> 1..9

<220> 
<223> The epitope from the first Ig extracellular domain of the CD22
      antigen, the farthest domain from the membrane

<400> 15
Glu Ser Thr Lys Asp Gly Lys Val Pro 
1               5                   

<210> 16
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> CD19-exon 2.1

<220> 
<221> misc_feature
<222> 1..20

<400> 16
caggcctggg aatccacatg                                                20


<210> 17
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> CD19-exon 14.1

<220> 
<221> misc_feature
<222> 1..20

<400> 17
agaacatgga taatcccgat                                                20


<210> 18
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> CD22-exon 3.2

<220> 
<221> misc_feature
<222> 1..20

<400> 18
tcaatgacag tggtcagctg                                                20


<210> 19
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> CD22-exon 9

<220> 
<221> misc_feature
<222> 1..20

<400> 19
caggtgtagt gggagacggg                                                20


<210> 20
<211> 117
<212> PRT
<213> Artificial Sequence


<220> 
<223> Sequence-based (and structure-assisted) CDR-grafting approach:
      Heavy chain

<400> 20
Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln 
1               5                   10                  15      
Thr Leu Ser Leu Thr Cys Ser Val Thr Gly Asp Ser Ile Thr Ser Gly 
            20                  25                  30          
Tyr Trp Asn Trp Ile Arg Lys Phe Pro Gly Asn Lys Leu Glu Trp Ile 
        35                  40                  45              
Gly Tyr Ile Ser Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser Leu Lys 
    50                  55                  60                  
Ser Arg Ile Thr Ile Ser Arg Asp Thr Ser Lys Asn Gln Tyr Ser Leu 
65                  70                  75                  80  
Lys Leu Ser Ser Val Thr Thr Glu Asp Thr Ala Thr Tyr Tyr Cys Ala 
                85                  90                  95      
Arg Tyr Pro Ser Pro Asp Ala Met Asn Tyr Trp Gly Gln Gly Thr Ser 
            100                 105                 110         
Val Thr Val Ser Ser 
        115         

<210> 21
<211> 113
<212> PRT
<213> Artificial Sequence


<220> 
<223> Sequence-based (and structure-assisted) CDR-grafting approach :
      Light chain

<400> 21
Asp Val Val Met Thr Gln Thr Pro Leu Thr Leu Pro Val Thr Leu Gly 
1               5                   10                  15      
Gln Pro Ala Ser Ile Ser Cys Lys Ser Ser Gln Ser Leu Leu Asp Ser 
            20                  25                  30          
Asp Gly Lys Thr Tyr Leu Asn Trp Leu Gln Gln Arg Pro Gly Gln Ser 
        35                  40                  45              
Pro Lys Arg Leu Ile Tyr Leu Val Ser Lys Leu Asp Ser Gly Val Pro 
    50                  55                  60                  
Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 
65                  70                  75                  80  
Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Trp Gln Gly 
                85                  90                  95      
Thr His Phe Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg 
            100                 105                 110         
Ala 
    

<210> 22
<211> 117
<212> PRT
<213> Artificial Sequence


<220> 
<223> The ‘De-murinenization’ approach, Heavy chain

<400> 22
Gln Val Gln Leu Gln Glu Ser Gly Pro Ser Leu Val Lys Pro Gly Gln 
1               5                   10                  15      
Thr Leu Ser Leu Thr Cys Ser Val Thr Gly Asp Ser Ile Thr Ser Gly 
            20                  25                  30          
Tyr Trp Asn Trp Ile Arg Gln Ser Pro Gly Asn Lys Leu Glu Tyr Met 
        35                  40                  45              
Gly Tyr Ile Ser Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Thr Leu Lys 
    50                  55                  60                  
Gly Arg Ile Ser Ile Thr Arg Asp Asn Ser Ser Ser Gln Tyr Tyr Leu 
65                  70                  75                  80  
Gln Leu Lys Ser Val Thr Ser Glu Asp Thr Ala Thr Phe Tyr Cys Ala 
                85                  90                  95      
Arg Tyr Pro Ser Pro Asp Ala Met Asn Tyr Trp Gly Gln Gly Thr Ser 
            100                 105                 110         
Val Thr Val Ser Ser 
        115         

<210> 23
<211> 113
<212> PRT
<213> Artificial Sequence


<220> 
<223> ‘De-murinenization’ approach, Light chain

<400> 23
Asp Val Val Met Thr Gln Thr Pro Leu Thr Leu Ser Val Thr Ile Gly 
1               5                   10                  15      
Gln Pro Ala Ser Ile Ser Cys Lys Ser Ser Gln Ser Leu Leu Asp Ser 
            20                  25                  30          
Asp Gly Lys Thr Tyr Leu Asn Trp Leu Leu Gln Arg Pro Gly Gln Ser 
        35                  40                  45              
Pro Lys Leu Leu Ile Tyr Leu Val Ser Lys Leu Asp Ser Gly Val Pro 
    50                  55                  60                  
Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 
65                  70                  75                  80  
Ser Arg Val Glu Ala Asp Asp Leu Gly Val Tyr Phe Cys Trp Gln Gly 
                85                  90                  95      
Thr His Phe Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg 
            100                 105                 110         
Ala 
    

