                         SEQUENCE LISTING

<110>  ISTITUTO ORTOPEDICO RIZZOLI
       ISTITUTO SUPERIORE DI SANITA'
 
<120>  Single-chain variable Fragment (scFv) able to recognize and bind 
       CD99 human protein.

<130>  61.I3257.12.WO.20

<160>  27    

<170>  PatentIn version 3.3

<210>  1
<211>  9
<212>  PRT
<213>  Artificial

<220>
<223>  CDR3 amino acid sequence of VH Chain

<400>  1

Ala Lys Ser His Lys Arg Phe Asp Tyr 
1               5                   


<210>  2
<211>  11
<212>  PRT
<213>  Artificial

<220>
<223>  CDR3 amino acid sequence of VL Chain

<400>  2

Asn Ser Ser Phe Pro Arg Thr Ser Ser Val Val 
1               5                   10      


<210>  3
<211>  10
<212>  PRT
<213>  Artificial

<220>
<223>  CDR1 amino acid sequence of VH Chain

<400>  3

Gly Phe Thr Phe Ser Ser Tyr Ala Met Ser 
1               5                   10  


<210>  4
<211>  9
<212>  PRT
<213>  Artificial

<220>
<223>  CDR2 amino acid sequence VH Chain

<400>  4

Ala Ile Ser Gly Ser Gly Gly Ser Thr 
1               5                   


<210>  5
<211>  11
<212>  PRT
<213>  Artificial

<220>
<223>  CDR1 amino acid sequence VL Chain

<400>  5

Gln Gly Asp Ser Leu Arg Ser Tyr Tyr Ala Ser 
1               5                   10      


<210>  6
<211>  7
<212>  PRT
<213>  Artificial

<220>
<223>  CDR2 amino acid sequence VL Chain

<400>  6

Gly Lys Asn Asn Arg Pro Ser 
1               5           


<210>  7
<211>  118
<212>  PRT
<213>  Artificial

<220>
<223>  VH Chain amino acid sequence

<400>  7

Met Ala Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Arg Pro 
1               5                   10                  15      


Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser 
            20                  25                  30          


Ser Tyr Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu 
        35                  40                  45              


Trp Val Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp 
    50                  55                  60                  


Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr 
65                  70                  75                  80  


Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr 
                85                  90                  95      


Tyr Cys Ala Lys Ser His Lys Arg Phe Asp Tyr Trp Gly Gln Gly Thr 
            100                 105                 110         


Leu Val Thr Val Ser Arg 
        115             


<210>  8
<211>  109
<212>  PRT
<213>  Artificial

<220>
<223>  VL Chain amino acid sequence

<400>  8

Ser Ser Glu Leu Thr Gln Asp Pro Ala Val Ser Val Ala Leu Gly Gln 
1               5                   10                  15      


Thr Val Arg Ile Thr Cys Gln Gly Asp Ser Leu Arg Ser Tyr Tyr Ala 
            20                  25                  30          


Ser Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Val Leu Val Ile Tyr 
        35                  40                  45              


Gly Lys Asn Asn Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser 
    50                  55                  60                  


Ser Ser Gly Asn Thr Ala Ser Leu Thr Ile Thr Gly Ala Gln Ala Glu 
65                  70                  75                  80  


Asp Glu Ala Asp Tyr Tyr Cys Asn Ser Ser Phe Pro Arg Thr Ser Ser 
                85                  90                  95      


Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly 
            100                 105                 


<210>  9
<211>  14
<212>  PRT
<213>  Artificial

<220>
<223>  Linker animo acid sequence

<400>  9

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 
1               5                   10                  


<210>  10
<211>  241
<212>  PRT
<213>  Artificial

<220>
<223>  scFv amino acid sequence

<400>  10

Met Ala Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Arg Pro 
1               5                   10                  15      


Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser 
            20                  25                  30          


Ser Tyr Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu 
        35                  40                  45              


Trp Val Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp 
    50                  55                  60                  


Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr 
65                  70                  75                  80  


Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr 
                85                  90                  95      


Tyr Cys Ala Lys Ser His Lys Arg Phe Asp Tyr Trp Gly Gln Gly Thr 
            100                 105                 110         


Leu Val Thr Val Ser Arg Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 
        115                 120                 125             


Gly Gly Gly Gly Ser Ser Glu Leu Thr Gln Asp Pro Ala Val Ser Val 
    130                 135                 140                 


Ala Leu Gly Gln Thr Val Arg Ile Thr Cys Gln Gly Asp Ser Leu Arg 
145                 150                 155                 160 


Ser Tyr Tyr Ala Ser Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Val 
                165                 170                 175     


Leu Val Ile Tyr Gly Lys Asn Asn Arg Pro Ser Gly Ile Pro Asp Arg 
            180                 185                 190         


Phe Ser Gly Ser Ser Ser Gly Asn Thr Ala Ser Leu Thr Ile Thr Gly 
        195                 200                 205             


Ala Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Asn Ser Ser Phe Pro 
    210                 215                 220                 


Arg Thr Ser Ser Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 
225                 230                 235                 240 


Gly 
    


<210>  11
<211>  27
<212>  DNA
<213>  Artificial

<220>
<223>  CDR3 nucleotide sequence of VH Chain

<400>  11
gcgaaatcgc ataagcgttt tgactac                                         27


<210>  12
<211>  33
<212>  DNA
<213>  Artificial

<220>
<223>  CDR3 nucleotide sequence of VL Chain

<400>  12
aactcctctt ttccccggac ttcttctgtg gta                                  33


<210>  13
<211>  30
<212>  DNA
<213>  Artificial

<220>
<223>  CDR1 nucleotide sequence of VH Chain

<400>  13
ggattcacct ttagcagcta tgccatgagc                                      30


<210>  14
<211>  27
<212>  DNA
<213>  Artificial

<220>
<223>  CDR2 nucleotide sequence of VH Chain

<400>  14
gctattagtg gtagtggtgg tagcaca                                         27


<210>  15
<211>  36
<212>  DNA
<213>  Artificial

<220>
<223>  CDR1 nucleotide sequence of VL Chain

<400>  15
tgccaaggag acagcctcag aagctattat gcaagc                               36


<210>  16
<211>  21
<212>  DNA
<213>  Artificial

<220>
<223>  CDR2 nucleotide sequence of VL Chain

<400>  16
ggtaaaaaca accggccctc a                                               21


<210>  17
<211>  354
<212>  DNA
<213>  Artificial

<220>
<223>  VH Chain nucleotide sequence

<400>  17
atggccgagg tgcagctggt ggagtctggg ggaggcttgg tacggcctgg ggggtccctg     60

agactctcct gtgcagcctc tggattcacc tttagcagct atgccatgag ctgggtccgc    120

caggctccag ggaaggggct ggagtgggtc tcagctatta gtggtagtgg tggtagcaca    180

tactacgcag actccgtgaa gggccggttc accatctcca gagacaattc caagaacacg    240

ctgtatctgc aaatgaacag cctgagagcc gaggacacgg ccgtatatta ctgtgcgaaa    300

tcgcataagc gttttgacta ctggggccag ggaaccctgg tcaccgtgtc gaga          354


<210>  18
<211>  327
<212>  DNA
<213>  Artificial

<220>
<223>  VL Chain nucleotide sequence

<400>  18
tcgtctgagc tgactcagga ccctgctgtg tctgtggcct tgggacagac agtcaggatc     60

acatgccaag gagacagcct cagaagctat tatgcaagct ggtaccagca gaagccagga    120

caggcccctg tacttgtcat ctatggtaaa aacaaccggc cctcagggat cccagaccga    180

ttctctggct ccagctcagg aaacacagct tccttgacca tcactggggc tcaggcggaa    240

gatgaggctg actattactg taactcctct tttccccgga cttcttctgt ggtattcggc    300

ggagggacca agctgaccgt cctaggc                                        327


<210>  19
<211>  42
<212>  DNA
<213>  Artificial

<220>
<223>  Linker nucleotide sequence

<400>  19
ggtggaggcg gttcaggcgg aggtggctct ggcggtggcg ga                        42


<210>  20
<211>  723
<212>  DNA
<213>  Artificial

<220>
<223>  scFv nucleotide sequence

<400>  20
atggccgagg tgcagctggt ggagtctggg ggaggcttgg tacggcctgg ggggtccctg     60

agactctcct gtgcagcctc tggattcacc tttagcagct atgccatgag ctgggtccgc    120

caggctccag ggaaggggct ggagtgggtc tcagctatta gtggtagtgg tggtagcaca    180

tactacgcag actccgtgaa gggccggttc accatctcca gagacaattc caagaacacg    240

ctgtatctgc aaatgaacag cctgagagcc gaggacacgg ccgtatatta ctgtgcgaaa    300

tcgcataagc gttttgacta ctggggccag ggaaccctgg tcaccgtgtc gagaggtgga    360

ggcggttcag gcggaggtgg ctctggcggt ggcggatcgt ctgagctgac tcaggaccct    420

gctgtgtctg tggccttggg acagacagtc aggatcacat gccaaggaga cagcctcaga    480

agctattatg caagctggta ccagcagaag ccaggacagg cccctgtact tgtcatctat    540

ggtaaaaaca accggccctc agggatccca gaccgattct ctggctccag ctcaggaaac    600

acagcttcct tgaccatcac tggggctcag gcggaagatg aggctgacta ttactgtaac    660

tcctcttttc cccggacttc ttctgtggta ttcggcggag ggaccaagct gaccgtccta    720

ggc                                                                  723


<210>  21
<211>  100
<212>  PRT
<213>  Artificial

<220>
<223>  Amino acid sequence of CD99 extracellular domain

<400>  21

Asp Gly Gly Phe Asp Leu Ser Asp Ala Leu Pro Asp Asn Glu Asn Lys 
1               5                   10                  15      


Lys Pro Thr Ala Ile Pro Lys Lys Pro Ser Ala Gly Asp Asp Phe Asp 
            20                  25                  30          


Leu Gly Asp Ala Val Val Asp Gly Glu Asn Asp Asp Pro Arg Pro Pro 
        35                  40                  45              


Asn Pro Pro Lys Pro Met Pro Asn Pro Asn Pro Asn His Pro Ser Ser 
    50                  55                  60                  


Ser Gly Ser Phe Ser Asp Ala Asp Leu Ala Asp Gly Val Ser Gly Gly 
65                  70                  75                  80  


Glu Gly Lys Gly Gly Ser Asp Gly Gly Gly Ser His Arg Lys Glu Gly 
                85                  90                  95      


Glu Glu Ala Asp 
            100 


<210>  22
<211>  300
<212>  DNA
<213>  Artificial

<220>
<223>  Nucleotide sequence of CD99 extracellular domain

<400>  22
gatggtggtt tcgatttatc cgatgccctt cctgacaatg agaacaagaa acccactgca     60

atccccaaga aacccagtgc tggggatgac tttgacttag gagatgctgt tgttgatgga    120

gaaaatgacg acccacgacc accgaaccca cccaaaccga tgccaaatcc aaaccccaac    180

caccctagtt cctccggtag cttttcagat gctgaccttg cggatggcgt ttcaggtgga    240

gaaggaaaag gaggcagtga tggtggaggc agccacagga aagaagggga agaggccgac    300


<210>  23
<211>  28
<212>  DNA
<213>  Artificial

<220>
<223>  CD99 For Primer

<400>  23
cgatggatcc gatggtggtt tcgattta                                        28


<210>  24
<211>  21
<212>  DNA
<213>  Artificial

<220>
<223>  CD99 Rev Primer

<400>  24
acatgtcgac gtcggcctct t                                               21


<210>  25
<211>  25
<212>  PRT
<213>  Artificial

<220>
<223>  CD99 amino acid sequence representing the epitope

<400>  25

Gly Asp Asp Phe Asp Leu Gly Asp Ala Val Val Asp Gly Glu Asn Asp 
1               5                   10                  15      


Asp Pro Arg Pro Pro Asn Pro Pro Lys 
            20                  25  


<210>  26
<211>  17
<212>  DNA
<213>  Artificial

<220>
<223>  fdseq1 Primer

<400>  26
ggattttctg tatgagg                                                    17


<210>  27
<211>  20
<212>  DNA
<213>  Artificial

<220>
<223>  pelBback

<400>  27
agccgctgga ttgttattac                                                 20


