                         SEQUENCE LISTING

<110>  Thomas Jefferson University
 
<120>  Anti-GUCY2C Chimeric Antigen Receptor Compositions and Methods

<130>  100051.19302

<150>  62/643,850
<151>  2018-03-16

<160>  15    

<170>  PatentIn version 3.5

<210>  1
<211>  1671
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic CAR-T containing human derived sequences


<220>
<221>  CDS
<222>  (1)..(1671)
<223>  5F9.28BBz CAR construct for human CAR-T cell expression

<220>
<221>  sig_peptide
<222>  (1)..(66)
<223>  hGMCSFRa Signal Peptide

<220>
<221>  misc_feature
<222>  (73)..(822)
<223>  5F9 anti-GUYC2C scFv

<220>
<221>  misc_feature
<222>  (829)..(1008)
<223>  CD8 hinge

<220>
<221>  misc_feature
<222>  (1009)..(1215)
<223>  CD28 TM

<220>
<221>  misc_feature
<222>  (1216)..(1332)
<223>  4-1BB intracellular - signalling domain

<220>
<221>  misc_feature
<222>  (1333)..(1671)
<223>  CD3zeta intracellular - signalling domain ITAM

<400>  1
atg ctg ctg ctc gtg aca tct ctg ctg ctg tgc gag ctg ccc cac ccc         48
Met Leu Leu Leu Val Thr Ser Leu Leu Leu Cys Glu Leu Pro His Pro           
1               5                   10                  15                

gcc ttt ctg ctg atc ccc gat atc gag atc gtg atg acc cag agc ccc         96
Ala Phe Leu Leu Ile Pro Asp Ile Glu Ile Val Met Thr Gln Ser Pro           
            20                  25                  30                    

gcc acc ctg agt gtg tct cca ggc gaa aga gcc acc ctg tcc tgc aga        144
Ala Thr Leu Ser Val Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg           
        35                  40                  45                        

gcc agc cag agc gtg tcc aga aac ctg gcc tgg tat cag cag aag ccc        192
Ala Ser Gln Ser Val Ser Arg Asn Leu Ala Trp Tyr Gln Gln Lys Pro           
    50                  55                  60                            

ggc cag gct ccc cgg ctg ctg atc tat ggc gcc agc aca aga gcc aca        240
Gly Gln Ala Pro Arg Leu Leu Ile Tyr Gly Ala Ser Thr Arg Ala Thr           
65                  70                  75                  80            

ggc atc ccc gcc aga ttt tcc ggc tct ggc agc ggc acc gag ttc acc        288
Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr           
                85                  90                  95                

ctg aca atc ggc agc ctg cag tcc gag gac ttc gcc gtg tac tac tgc        336
Leu Thr Ile Gly Ser Leu Gln Ser Glu Asp Phe Ala Val Tyr Tyr Cys           
            100                 105                 110                   

cag cag tac aag acc tgg ccc cgg acc ttt ggc cag ggc acc aac gtg        384
Gln Gln Tyr Lys Thr Trp Pro Arg Thr Phe Gly Gln Gly Thr Asn Val           
        115                 120                 125                       

gaa atc aag gcc tct ggc ggc gga gga tct ggg gga ggc gga agc gga        432
Glu Ile Lys Ala Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly           
    130                 135                 140                           

ggc gga gga agt ggc gga ggg gga tct gaa ctg cag gtg cag ctg cag        480
Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Leu Gln Val Gln Leu Gln           
145                 150                 155                 160           

cag tgg gga gcc gga ctg ctg aag cct agc gag aca ctg agc ctg acc        528
Gln Trp Gly Ala Gly Leu Leu Lys Pro Ser Glu Thr Leu Ser Leu Thr           
                165                 170                 175               

tgc gcc gtg ttc ggc ggc agc ttc agc ggc tac tac tgg tcc tgg atc        576
Cys Ala Val Phe Gly Gly Ser Phe Ser Gly Tyr Tyr Trp Ser Trp Ile           
            180                 185                 190                   

aga cag ccc cct ggc aag ggc ctg gaa tgg atc ggc gag atc aac cac        624
Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile Gly Glu Ile Asn His           
        195                 200                 205                       

cgg ggc aac acc aac gac aac ccc agc ctg aag tcc aga gtg acc atc        672
Arg Gly Asn Thr Asn Asp Asn Pro Ser Leu Lys Ser Arg Val Thr Ile           
    210                 215                 220                           

agc gtg gac acc agc aag aac cag ttc gcc ctg aag ctg agc agc gtg        720
Ser Val Asp Thr Ser Lys Asn Gln Phe Ala Leu Lys Leu Ser Ser Val           
225                 230                 235                 240           

aca gcc gcc gat acc gcc gtg tat tat tgc gcc aga gag cgg ggc tac        768
Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala Arg Glu Arg Gly Tyr           
                245                 250                 255               

acc tac ggc aac ttc gac cac tgg ggc cag gga acc ctc gtg acc gtg        816
Thr Tyr Gly Asn Phe Asp His Trp Gly Gln Gly Thr Leu Val Thr Val           
            260                 265                 270                   

tct agc gag ctc ctg agc aac agc atc atg tac ttc agc cac ttc gtg        864
Ser Ser Glu Leu Leu Ser Asn Ser Ile Met Tyr Phe Ser His Phe Val           
        275                 280                 285                       

ccc gtg ttt ctg ccc gcc aag cct acc aca acc cct gcc cct aga cct        912
Pro Val Phe Leu Pro Ala Lys Pro Thr Thr Thr Pro Ala Pro Arg Pro           
    290                 295                 300                           

cct acc cca gcc cct aca atc gcc tcc cag cct ctg tct ctg agg ccc        960
Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro           
305                 310                 315                 320           

gag gct tgt aga cct gct gct ggc gga gcc gtg cac acc aga gga ctg       1008
Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu           
                325                 330                 335               

ttt tgg gtg ctg gtg gtc gtg ggc gga gtg ctg gcc tgt tac agc ctg       1056
Phe Trp Val Leu Val Val Val Gly Gly Val Leu Ala Cys Tyr Ser Leu           
            340                 345                 350                   

ctc gtg act gtg gcc ttc atc atc ttt tgg gtg cgc agc aag cgg agc       1104
Leu Val Thr Val Ala Phe Ile Ile Phe Trp Val Arg Ser Lys Arg Ser           
        355                 360                 365                       

aga ctg ctg cac agc gac tac atg aac atg acc ccc aga cgg cca ggc       1152
Arg Leu Leu His Ser Asp Tyr Met Asn Met Thr Pro Arg Arg Pro Gly           
    370                 375                 380                           

ccc acc aga aag cac tac cag cct tac gcc cct ccc cgg gat ttc gcc       1200
Pro Thr Arg Lys His Tyr Gln Pro Tyr Ala Pro Pro Arg Asp Phe Ala           
385                 390                 395                 400           

gcc tac aga agc ggc aga aag aag ctg ctg tac atc ttc aag cag ccc       1248
Ala Tyr Arg Ser Gly Arg Lys Lys Leu Leu Tyr Ile Phe Lys Gln Pro           
                405                 410                 415               

ttc atg cgg ccc gtg cag acc acc cag gaa gag gac ggc tgc tcc tgc       1296
Phe Met Arg Pro Val Gln Thr Thr Gln Glu Glu Asp Gly Cys Ser Cys           
            420                 425                 430                   

aga ttc ccc gag gaa gaa gaa ggc ggc tgc gag ctg aga gtg aag ttc       1344
Arg Phe Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu Arg Val Lys Phe           
        435                 440                 445                       

agc aga tcc gcc gac gcc cct gcc tat cag cag ggc cag aac cag ctg       1392
Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu           
    450                 455                 460                           

tac aac gag ctg aac ctg ggc aga cgg gaa gag tac gac gtg ctg gac       1440
Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp           
465                 470                 475                 480           

aag cgg aga ggc agg gac cct gag atg ggc gga aag ccc cag aga aga       1488
Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Gln Arg Arg           
                485                 490                 495               

aag aac ccc cag gaa ggc ctg tat aac gaa ctg cag aaa gac aag atg       1536
Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met           
            500                 505                 510                   

gcc gag gcc tac agc gag atc gga atg aag ggc gag cgg aga aga ggc       1584
Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly           
        515                 520                 525                       

aag gga cac gac ggc ctg tac cag gga ctg agc acc gcc acc aag gac       1632
Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp           
    530                 535                 540                           

acc tac gac gcc ctg cac atg cag gcc ctg ccc ccc aga                   1671
Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg                       
545                 550                 555                               


<210>  2
<211>  557
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic Construct

<400>  2

Met Leu Leu Leu Val Thr Ser Leu Leu Leu Cys Glu Leu Pro His Pro 
1               5                   10                  15      


Ala Phe Leu Leu Ile Pro Asp Ile Glu Ile Val Met Thr Gln Ser Pro 
            20                  25                  30          


Ala Thr Leu Ser Val Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg 
        35                  40                  45              


Ala Ser Gln Ser Val Ser Arg Asn Leu Ala Trp Tyr Gln Gln Lys Pro 
    50                  55                  60                  


Gly Gln Ala Pro Arg Leu Leu Ile Tyr Gly Ala Ser Thr Arg Ala Thr 
65                  70                  75                  80  


Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr 
                85                  90                  95      


Leu Thr Ile Gly Ser Leu Gln Ser Glu Asp Phe Ala Val Tyr Tyr Cys 
            100                 105                 110         


Gln Gln Tyr Lys Thr Trp Pro Arg Thr Phe Gly Gln Gly Thr Asn Val 
        115                 120                 125             


Glu Ile Lys Ala Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 
    130                 135                 140                 


Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Leu Gln Val Gln Leu Gln 
145                 150                 155                 160 


Gln Trp Gly Ala Gly Leu Leu Lys Pro Ser Glu Thr Leu Ser Leu Thr 
                165                 170                 175     


Cys Ala Val Phe Gly Gly Ser Phe Ser Gly Tyr Tyr Trp Ser Trp Ile 
            180                 185                 190         


Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile Gly Glu Ile Asn His 
        195                 200                 205             


Arg Gly Asn Thr Asn Asp Asn Pro Ser Leu Lys Ser Arg Val Thr Ile 
    210                 215                 220                 


Ser Val Asp Thr Ser Lys Asn Gln Phe Ala Leu Lys Leu Ser Ser Val 
225                 230                 235                 240 


Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala Arg Glu Arg Gly Tyr 
                245                 250                 255     


Thr Tyr Gly Asn Phe Asp His Trp Gly Gln Gly Thr Leu Val Thr Val 
            260                 265                 270         


Ser Ser Glu Leu Leu Ser Asn Ser Ile Met Tyr Phe Ser His Phe Val 
        275                 280                 285             


Pro Val Phe Leu Pro Ala Lys Pro Thr Thr Thr Pro Ala Pro Arg Pro 
    290                 295                 300                 


Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro 
305                 310                 315                 320 


Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu 
                325                 330                 335     


Phe Trp Val Leu Val Val Val Gly Gly Val Leu Ala Cys Tyr Ser Leu 
            340                 345                 350         


Leu Val Thr Val Ala Phe Ile Ile Phe Trp Val Arg Ser Lys Arg Ser 
        355                 360                 365             


Arg Leu Leu His Ser Asp Tyr Met Asn Met Thr Pro Arg Arg Pro Gly 
    370                 375                 380                 


Pro Thr Arg Lys His Tyr Gln Pro Tyr Ala Pro Pro Arg Asp Phe Ala 
385                 390                 395                 400 


Ala Tyr Arg Ser Gly Arg Lys Lys Leu Leu Tyr Ile Phe Lys Gln Pro 
                405                 410                 415     


Phe Met Arg Pro Val Gln Thr Thr Gln Glu Glu Asp Gly Cys Ser Cys 
            420                 425                 430         


Arg Phe Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu Arg Val Lys Phe 
        435                 440                 445             


Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu 
    450                 455                 460                 


Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp 
465                 470                 475                 480 


Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Gln Arg Arg 
                485                 490                 495     


Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met 
            500                 505                 510         


Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly 
        515                 520                 525             


Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp 
    530                 535                 540                 


Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg 
545                 550                 555         


<210>  3
<211>  5
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic linker sequence for scFV

<400>  3

Gly Gly Gly Gly Ser 
1               5   


<210>  4
<211>  10
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic linker sequence for scFV

<400>  4

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


<210>  5
<211>  15
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic linker sequence for scFV

<400>  5

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 
1               5                   10                  15  


<210>  6
<211>  20
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic linker sequence for scFV

<400>  6

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 
1               5                   10                  15      


Gly Gly Gly Ser 
            20  


<210>  7
<211>  25
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic linker sequence for scFV

<400>  7

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 
1               5                   10                  15      


Gly Gly Gly Ser Gly Gly Gly Gly Ser 
            20                  25  


<210>  8
<211>  240
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic construct scFv VL-L2-VH


<220>
<221>  MISC_FEATURE
<222>  (1)..(109)
<223>  5F9 Variable Light Chain fragment

<220>
<221>  MISC_FEATURE
<222>  (110)..(119)
<223>  Linker G4S-2

<220>
<221>  MISC_FEATURE
<222>  (120)..(240)
<223>  5F9 Variable Heavy Chain fragment

<400>  8

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


Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Arg Asn 
            20                  25                  30          


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


Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 
    50                  55                  60                  


Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Gly Ser Leu Gln Ser 
65                  70                  75                  80  


Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Lys Thr Trp Pro Arg 
                85                  90                  95      


Thr Phe Gly Gln Gly Thr Asn Val Glu Ile Lys Ala Ser Gly Gly Gly 
            100                 105                 110         


Gly Ser Gly Gly Gly Gly Ser Glu Leu Gln Val Gln Leu Gln Gln Trp 
        115                 120                 125             


Gly Ala Gly Leu Leu Lys Pro Ser Glu Thr Leu Ser Leu Thr Cys Ala 
    130                 135                 140                 


Val Phe Gly Gly Ser Phe Ser Gly Tyr Tyr Trp Ser Trp Ile Arg Gln 
145                 150                 155                 160 


Pro Pro Gly Lys Gly Leu Glu Trp Ile Gly Glu Ile Asn His Arg Gly 
                165                 170                 175     


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


Asp Thr Ser Lys Asn Gln Phe Ala Leu Lys Leu Ser Ser Val Thr Ala 
        195                 200                 205             


Ala Asp Thr Ala Val Tyr Tyr Cys Ala Arg Glu Arg Gly Tyr Thr Tyr 
    210                 215                 220                 


Gly Asn Phe Asp His Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 
225                 230                 235                 240 


<210>  9
<211>  245
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic construct scFv VL-L3-VH


<220>
<221>  MISC_FEATURE
<222>  (1)..(109)
<223>  5F9 Variable Light Chain fragment

<220>
<221>  MISC_FEATURE
<222>  (110)..(124)
<223>  Linker G4S-3

<220>
<221>  MISC_FEATURE
<222>  (125)..(245)
<223>  5F9 Variable Heavy Chain fragment

<400>  9

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


Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Arg Asn 
            20                  25                  30          


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


Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 
    50                  55                  60                  


Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Gly Ser Leu Gln Ser 
65                  70                  75                  80  


Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Lys Thr Trp Pro Arg 
                85                  90                  95      


Thr Phe Gly Gln Gly Thr Asn Val Glu Ile Lys Ala Ser Gly Gly Gly 
            100                 105                 110         


Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Leu Gln Val 
        115                 120                 125             


Gln Leu Gln Gln Trp Gly Ala Gly Leu Leu Lys Pro Ser Glu Thr Leu 
    130                 135                 140                 


Ser Leu Thr Cys Ala Val Phe Gly Gly Ser Phe Ser Gly Tyr Tyr Trp 
145                 150                 155                 160 


Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile Gly Glu 
                165                 170                 175     


Ile Asn His Arg Gly Asn Thr Asn Asp Asn Pro Ser Leu Lys Ser Arg 
            180                 185                 190         


Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe Ala Leu Lys Leu 
        195                 200                 205             


Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala Arg Glu 
    210                 215                 220                 


Arg Gly Tyr Thr Tyr Gly Asn Phe Asp His Trp Gly Gln Gly Thr Leu 
225                 230                 235                 240 


Val Thr Val Ser Ser 
                245 


<210>  10
<211>  250
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic construct scFv VL-L4-VH


<220>
<221>  MISC_FEATURE
<222>  (1)..(109)
<223>  5F9 Variable Light Chain fragment

<220>
<221>  MISC_FEATURE
<222>  (110)..(129)
<223>  Linker G4S-4

<220>
<221>  MISC_FEATURE
<222>  (130)..(250)
<223>  5F9 Variable Heavy Chain fragment

<400>  10

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


Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Arg Asn 
            20                  25                  30          


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


Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 
    50                  55                  60                  


Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Gly Ser Leu Gln Ser 
65                  70                  75                  80  


Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Lys Thr Trp Pro Arg 
                85                  90                  95      


Thr Phe Gly Gln Gly Thr Asn Val Glu Ile Lys Ala Ser Gly Gly Gly 
            100                 105                 110         


Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 
        115                 120                 125             


Ser Glu Leu Gln Val Gln Leu Gln Gln Trp Gly Ala Gly Leu Leu Lys 
    130                 135                 140                 


Pro Ser Glu Thr Leu Ser Leu Thr Cys Ala Val Phe Gly Gly Ser Phe 
145                 150                 155                 160 


Ser Gly Tyr Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu 
                165                 170                 175     


Glu Trp Ile Gly Glu Ile Asn His Arg Gly Asn Thr Asn Asp Asn Pro 
            180                 185                 190         


Ser Leu Lys Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln 
        195                 200                 205             


Phe Ala Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr 
    210                 215                 220                 


Tyr Cys Ala Arg Glu Arg Gly Tyr Thr Tyr Gly Asn Phe Asp His Trp 
225                 230                 235                 240 


Gly Gln Gly Thr Leu Val Thr Val Ser Ser 
                245                 250 


<210>  11
<211>  255
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic construct scFv VL-L5-VH


<220>
<221>  MISC_FEATURE
<222>  (1)..(109)
<223>  5F9 Variable Light Chain fragment

<220>
<221>  MISC_FEATURE
<222>  (110)..(134)
<223>  Linker G4S-5

<220>
<221>  MISC_FEATURE
<222>  (135)..(255)
<223>  5F9 Variable Heavy Chain fragment

<400>  11

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


Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Arg Asn 
            20                  25                  30          


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


Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 
    50                  55                  60                  


Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Gly Ser Leu Gln Ser 
65                  70                  75                  80  


Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Lys Thr Trp Pro Arg 
                85                  90                  95      


Thr Phe Gly Gln Gly Thr Asn Val Glu Ile Lys Ala Ser Gly Gly Gly 
            100                 105                 110         


Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 
        115                 120                 125             


Ser Gly Gly Gly Gly Ser Glu Leu Gln Val Gln Leu Gln Gln Trp Gly 
    130                 135                 140                 


Ala Gly Leu Leu Lys Pro Ser Glu Thr Leu Ser Leu Thr Cys Ala Val 
145                 150                 155                 160 


Phe Gly Gly Ser Phe Ser Gly Tyr Tyr Trp Ser Trp Ile Arg Gln Pro 
                165                 170                 175     


Pro Gly Lys Gly Leu Glu Trp Ile Gly Glu Ile Asn His Arg Gly Asn 
            180                 185                 190         


Thr Asn Asp Asn Pro Ser Leu Lys Ser Arg Val Thr Ile Ser Val Asp 
        195                 200                 205             


Thr Ser Lys Asn Gln Phe Ala Leu Lys Leu Ser Ser Val Thr Ala Ala 
    210                 215                 220                 


Asp Thr Ala Val Tyr Tyr Cys Ala Arg Glu Arg Gly Tyr Thr Tyr Gly 
225                 230                 235                 240 


Asn Phe Asp His Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 
                245                 250                 255 


<210>  12
<211>  240
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic construct scFv VH-L2-VL


<220>
<221>  MISC_FEATURE
<222>  (1)..(121)
<223>  5F9 Variable Heavy Chain fragment

<220>
<221>  MISC_FEATURE
<222>  (122)..(131)
<223>  Linker G4S-2

<220>
<221>  MISC_FEATURE
<222>  (132)..(240)
<223>  5F9 Variable Light Chain fragment

<400>  12

Glu Leu Gln Val Gln Leu Gln Gln Trp Gly Ala Gly Leu Leu Lys Pro 
1               5                   10                  15      


Ser Glu Thr Leu Ser Leu Thr Cys Ala Val Phe Gly Gly Ser Phe Ser 
            20                  25                  30          


Gly Tyr Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 
        35                  40                  45              


Trp Ile Gly Glu Ile Asn His Arg Gly Asn Thr Asn Asp Asn Pro Ser 
    50                  55                  60                  


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


Ala Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr 
                85                  90                  95      


Cys Ala Arg Glu Arg Gly Tyr Thr Tyr Gly Asn Phe Asp His Trp Gly 
            100                 105                 110         


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


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


Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val 
145                 150                 155                 160 


Ser Arg Asn Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg 
                165                 170                 175     


Leu Leu Ile Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg 
            180                 185                 190         


Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Gly Ser 
        195                 200                 205             


Leu Gln Ser Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Lys Thr 
    210                 215                 220                 


Trp Pro Arg Thr Phe Gly Gln Gly Thr Asn Val Glu Ile Lys Ala Ser 
225                 230                 235                 240 


<210>  13
<211>  245
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic construct scFv VH-L3-VL


<220>
<221>  MISC_FEATURE
<222>  (1)..(121)
<223>  5F9 Variable Heavy Chain fragment

<220>
<221>  MISC_FEATURE
<222>  (122)..(136)
<223>  Linker G4S-3

<220>
<221>  MISC_FEATURE
<222>  (137)..(245)
<223>  5F9 Variable Light Chain fragment

<400>  13

Glu Leu Gln Val Gln Leu Gln Gln Trp Gly Ala Gly Leu Leu Lys Pro 
1               5                   10                  15      


Ser Glu Thr Leu Ser Leu Thr Cys Ala Val Phe Gly Gly Ser Phe Ser 
            20                  25                  30          


Gly Tyr Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 
        35                  40                  45              


Trp Ile Gly Glu Ile Asn His Arg Gly Asn Thr Asn Asp Asn Pro Ser 
    50                  55                  60                  


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


Ala Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr 
                85                  90                  95      


Cys Ala Arg Glu Arg Gly Tyr Thr Tyr Gly Asn Phe Asp His Trp Gly 
            100                 105                 110         


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


Gly Gly Ser Gly Gly Gly Gly Ser Glu Ile Val Met Thr Gln Ser Pro 
    130                 135                 140                 


Ala Thr Leu Ser Val Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg 
145                 150                 155                 160 


Ala Ser Gln Ser Val Ser Arg Asn Leu Ala Trp Tyr Gln Gln Lys Pro 
                165                 170                 175     


Gly Gln Ala Pro Arg Leu Leu Ile Tyr Gly Ala Ser Thr Arg Ala Thr 
            180                 185                 190         


Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr 
        195                 200                 205             


Leu Thr Ile Gly Ser Leu Gln Ser Glu Asp Phe Ala Val Tyr Tyr Cys 
    210                 215                 220                 


Gln Gln Tyr Lys Thr Trp Pro Arg Thr Phe Gly Gln Gly Thr Asn Val 
225                 230                 235                 240 


Glu Ile Lys Ala Ser 
                245 


<210>  14
<211>  250
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic construct scFv VH-L4-VL


<220>
<221>  MISC_FEATURE
<222>  (1)..(121)
<223>  5F9 Variable Heavy Chain fragment

<220>
<221>  MISC_FEATURE
<222>  (122)..(141)
<223>  Linker G4S-4

<220>
<221>  MISC_FEATURE
<222>  (142)..(250)
<223>  5F9 Variable Light Chain fragment

<400>  14

Glu Leu Gln Val Gln Leu Gln Gln Trp Gly Ala Gly Leu Leu Lys Pro 
1               5                   10                  15      


Ser Glu Thr Leu Ser Leu Thr Cys Ala Val Phe Gly Gly Ser Phe Ser 
            20                  25                  30          


Gly Tyr Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 
        35                  40                  45              


Trp Ile Gly Glu Ile Asn His Arg Gly Asn Thr Asn Asp Asn Pro Ser 
    50                  55                  60                  


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


Ala Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr 
                85                  90                  95      


Cys Ala Arg Glu Arg Gly Tyr Thr Tyr Gly Asn Phe Asp His Trp Gly 
            100                 105                 110         


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


Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Ile Val 
    130                 135                 140                 


Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly Glu Arg Ala 
145                 150                 155                 160 


Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Arg Asn Leu Ala Trp 
                165                 170                 175     


Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr Gly Ala 
            180                 185                 190         


Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser 
        195                 200                 205             


Gly Thr Glu Phe Thr Leu Thr Ile Gly Ser Leu Gln Ser Glu Asp Phe 
    210                 215                 220                 


Ala Val Tyr Tyr Cys Gln Gln Tyr Lys Thr Trp Pro Arg Thr Phe Gly 
225                 230                 235                 240 


Gln Gly Thr Asn Val Glu Ile Lys Ala Ser 
                245                 250 


<210>  15
<211>  255
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic construct scFv VH-L5-VL


<220>
<221>  MISC_FEATURE
<222>  (1)..(121)
<223>  5F9 Variable Heavy Chain fragment

<220>
<221>  MISC_FEATURE
<222>  (122)..(146)
<223>  Linker G4S-5

<220>
<221>  MISC_FEATURE
<222>  (147)..(255)
<223>  5F9 Variable Light Chain fragment

<400>  15

Glu Leu Gln Val Gln Leu Gln Gln Trp Gly Ala Gly Leu Leu Lys Pro 
1               5                   10                  15      


Ser Glu Thr Leu Ser Leu Thr Cys Ala Val Phe Gly Gly Ser Phe Ser 
            20                  25                  30          


Gly Tyr Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 
        35                  40                  45              


Trp Ile Gly Glu Ile Asn His Arg Gly Asn Thr Asn Asp Asn Pro Ser 
    50                  55                  60                  


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


Ala Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr 
                85                  90                  95      


Cys Ala Arg Glu Arg Gly Tyr Thr Tyr Gly Asn Phe Asp His Trp Gly 
            100                 105                 110         


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


Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly 
    130                 135                 140                 


Gly Ser Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser 
145                 150                 155                 160 


Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser 
                165                 170                 175     


Arg Asn Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu 
            180                 185                 190         


Leu Ile Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe 
        195                 200                 205             


Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Gly Ser Leu 
    210                 215                 220                 


Gln Ser Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Lys Thr Trp 
225                 230                 235                 240 


Pro Arg Thr Phe Gly Gln Gly Thr Asn Val Glu Ile Lys Ala Ser 
                245                 250                 255 


