                               SEQUENCE LISTING

<110> THE BRIGHAM AND WOMEN'S HOSPITAL, INC.
 
<120> METHODS TO MANIPULATE ALPHA-FETOPROTEIN (AFP)

<130> 043214-081612-PCT

<140> PCT/US15/26860
<141> 2015-04-21

<150> 62/101,539
<151> 2015-01-09

<150> 61/984,252
<151> 2014-04-25

<160> 31    

<170> PatentIn version 3.5

<210> 1
<211> 365
<212> PRT
<213> Homo sapiens

<400> 1
Met Gly Val Pro Arg Pro Gln Pro Trp Ala Leu Gly Leu Leu Leu Phe 
1               5                   10                  15      


Leu Leu Pro Gly Ser Leu Gly Ala Glu Ser His Leu Ser Leu Leu Tyr 
            20                  25                  30          


His Leu Thr Ala Val Ser Ser Pro Ala Pro Gly Thr Pro Ala Phe Trp 
        35                  40                  45              


Val Ser Gly Trp Leu Gly Pro Gln Gln Tyr Leu Ser Tyr Asn Ser Leu 
    50                  55                  60                  


Arg Gly Glu Ala Glu Pro Cys Gly Ala Trp Val Trp Glu Asn Gln Val 
65                  70                  75                  80  


Ser Trp Tyr Trp Glu Lys Glu Thr Thr Asp Leu Arg Ile Lys Glu Lys 
                85                  90                  95      


Leu Phe Leu Glu Ala Phe Lys Ala Leu Gly Gly Lys Gly Pro Tyr Thr 
            100                 105                 110         


Leu Gln Gly Leu Leu Gly Cys Glu Leu Gly Pro Asp Asn Thr Ser Val 
        115                 120                 125             


Pro Thr Ala Lys Phe Ala Leu Asn Gly Glu Glu Phe Met Asn Phe Asp 
    130                 135                 140                 


Leu Lys Gln Gly Thr Trp Gly Gly Asp Trp Pro Glu Ala Leu Ala Ile 
145                 150                 155                 160 


Ser Gln Arg Trp Gln Gln Gln Asp Lys Ala Ala Asn Lys Glu Leu Thr 
                165                 170                 175     


Phe Leu Leu Phe Ser Cys Pro His Arg Leu Arg Glu His Leu Glu Arg 
            180                 185                 190         


Gly Arg Gly Asn Leu Glu Trp Lys Glu Pro Pro Ser Met Arg Leu Lys 
        195                 200                 205             


Ala Arg Pro Ser Ser Pro Gly Phe Ser Val Leu Thr Cys Ser Ala Phe 
    210                 215                 220                 


Ser Phe Tyr Pro Pro Glu Leu Gln Leu Arg Phe Leu Arg Asn Gly Leu 
225                 230                 235                 240 


Ala Ala Gly Thr Gly Gln Gly Asp Phe Gly Pro Asn Ser Asp Gly Ser 
                245                 250                 255     


Phe His Ala Ser Ser Ser Leu Thr Val Lys Ser Gly Asp Glu His His 
            260                 265                 270         


Tyr Cys Cys Ile Val Gln His Ala Gly Leu Ala Gln Pro Leu Arg Val 
        275                 280                 285             


Glu Leu Glu Ser Pro Ala Lys Ser Ser Val Leu Val Val Gly Ile Val 
    290                 295                 300                 


Ile Gly Val Leu Leu Leu Thr Ala Ala Ala Val Gly Gly Ala Leu Leu 
305                 310                 315                 320 


Trp Arg Arg Met Arg Ser Gly Leu Pro Ala Pro Trp Ile Ser Leu Arg 
                325                 330                 335     


Gly Asp Asp Thr Gly Val Leu Leu Pro Thr Pro Gly Glu Ala Gln Asp 
            340                 345                 350         


Ala Asp Leu Lys Asp Val Asn Val Ile Pro Ala Thr Ala 
        355                 360                 365 


<210> 2
<211> 119
<212> PRT
<213> Homo sapiens

<400> 2
Met Ser Arg Ser Val Ala Leu Ala Val Leu Ala Leu Leu Ser Leu Ser 
1               5                   10                  15      


Gly Leu Glu Ala Ile Gln Arg Thr Pro Lys Ile Gln Val Tyr Ser Arg 
            20                  25                  30          


His Pro Ala Glu Asn Gly Lys Ser Asn Phe Leu Asn Cys Tyr Val Ser 
        35                  40                  45              


Gly Phe His Pro Ser Asp Ile Glu Val Asp Leu Leu Lys Asn Gly Glu 
    50                  55                  60                  


Arg Ile Glu Lys Val Glu His Ser Asp Leu Ser Phe Ser Lys Asp Trp 
65                  70                  75                  80  


Ser Phe Tyr Leu Leu Tyr Tyr Thr Glu Phe Thr Pro Thr Glu Lys Asp 
                85                  90                  95      


Glu Tyr Ala Cys Arg Val Asn His Val Thr Leu Ser Gln Pro Lys Ile 
            100                 105                 110         


Val Lys Trp Asp Arg Asp Met 
        115                 


<210> 3
<211> 609
<212> PRT
<213> Homo sapiens

<400> 3
Met Lys Trp Val Glu Ser Ile Phe Leu Ile Phe Leu Leu Asn Phe Thr 
1               5                   10                  15      


Glu Ser Arg Thr Leu His Arg Asn Glu Tyr Gly Ile Ala Ser Ile Leu 
            20                  25                  30          


Asp Ser Tyr Gln Cys Thr Ala Glu Ile Ser Leu Ala Asp Leu Ala Thr 
        35                  40                  45              


Ile Phe Phe Ala Gln Phe Val Gln Glu Ala Thr Tyr Lys Glu Val Ser 
    50                  55                  60                  


Lys Met Val Lys Asp Ala Leu Thr Ala Ile Glu Lys Pro Thr Gly Asp 
65                  70                  75                  80  


Glu Gln Ser Ser Gly Cys Leu Glu Asn Gln Leu Pro Ala Phe Leu Glu 
                85                  90                  95      


Glu Leu Cys His Glu Lys Glu Ile Leu Glu Lys Tyr Gly His Ser Asp 
            100                 105                 110         


Cys Cys Ser Gln Ser Glu Glu Gly Arg His Asn Cys Phe Leu Ala His 
        115                 120                 125             


Lys Lys Pro Thr Pro Ala Ser Ile Pro Leu Phe Gln Val Pro Glu Pro 
    130                 135                 140                 


Val Thr Ser Cys Glu Ala Tyr Glu Glu Asp Arg Glu Thr Phe Met Asn 
145                 150                 155                 160 


Lys Phe Ile Tyr Glu Ile Ala Arg Arg His Pro Phe Leu Tyr Ala Pro 
                165                 170                 175     


Thr Ile Leu Leu Trp Ala Ala Arg Tyr Asp Lys Ile Ile Pro Ser Cys 
            180                 185                 190         


Cys Lys Ala Glu Asn Ala Val Glu Cys Phe Gln Thr Lys Ala Ala Thr 
        195                 200                 205             


Val Thr Lys Glu Leu Arg Glu Ser Ser Leu Leu Asn Gln His Ala Cys 
    210                 215                 220                 


Ala Val Met Lys Asn Phe Gly Thr Arg Thr Phe Gln Ala Ile Thr Val 
225                 230                 235                 240 


Thr Lys Leu Ser Gln Lys Phe Thr Lys Val Asn Phe Thr Glu Ile Gln 
                245                 250                 255     


Lys Leu Val Leu Asp Val Ala His Val His Glu His Cys Cys Arg Gly 
            260                 265                 270         


Asp Val Leu Asp Cys Leu Gln Asp Gly Glu Lys Ile Met Ser Tyr Ile 
        275                 280                 285             


Cys Ser Gln Gln Asp Thr Leu Ser Asn Lys Ile Thr Glu Cys Cys Lys 
    290                 295                 300                 


Leu Thr Thr Leu Glu Arg Gly Gln Cys Ile Ile His Ala Glu Asn Asp 
305                 310                 315                 320 


Glu Lys Pro Glu Gly Leu Ser Pro Asn Leu Asn Arg Phe Leu Gly Asp 
                325                 330                 335     


Arg Asp Phe Asn Gln Phe Ser Ser Gly Glu Lys Asn Ile Phe Leu Ala 
            340                 345                 350         


Ser Phe Val His Glu Tyr Ser Arg Arg His Pro Gln Leu Ala Val Ser 
        355                 360                 365             


Val Ile Leu Arg Val Ala Lys Gly Tyr Gln Glu Leu Leu Glu Lys Cys 
    370                 375                 380                 


Phe Gln Thr Glu Asn Pro Leu Glu Cys Gln Asp Lys Gly Glu Glu Glu 
385                 390                 395                 400 


Leu Gln Lys Tyr Ile Gln Glu Ser Gln Ala Leu Ala Lys Arg Ser Cys 
                405                 410                 415     


Gly Leu Phe Gln Lys Leu Gly Glu Tyr Tyr Leu Gln Asn Ala Phe Leu 
            420                 425                 430         


Val Ala Tyr Thr Lys Lys Ala Pro Gln Leu Thr Ser Ser Glu Leu Met 
        435                 440                 445             


Ala Ile Thr Arg Lys Met Ala Ala Thr Ala Ala Thr Cys Cys Gln Leu 
    450                 455                 460                 


Ser Glu Asp Lys Leu Leu Ala Cys Gly Glu Gly Ala Ala Asp Ile Ile 
465                 470                 475                 480 


Ile Gly His Leu Cys Ile Arg His Glu Met Thr Pro Val Asn Pro Gly 
                485                 490                 495     


Val Gly Gln Cys Cys Thr Ser Ser Tyr Ala Asn Arg Arg Pro Cys Phe 
            500                 505                 510         


Ser Ser Leu Val Val Asp Glu Thr Tyr Val Pro Pro Ala Phe Ser Asp 
        515                 520                 525             


Asp Lys Phe Ile Phe His Lys Asp Leu Cys Gln Ala Gln Gly Val Ala 
    530                 535                 540                 


Leu Gln Thr Met Lys Gln Glu Phe Leu Ile Asn Leu Val Lys Gln Lys 
545                 550                 555                 560 


Pro Gln Ile Thr Glu Glu Gln Leu Glu Ala Val Ile Ala Asp Phe Ser 
                565                 570                 575     


Gly Leu Leu Glu Lys Cys Cys Gln Gly Gln Glu Gln Glu Val Cys Phe 
            580                 585                 590         


Ala Glu Glu Gly Gln Lys Leu Ile Ser Lys Thr Arg Ala Ala Leu Gly 
        595                 600                 605             


Val 
    


<210> 4
<211> 575
<212> PRT
<213> Artificial Sequence

<220>
<223> Description of Artificial Sequence: Synthetic
      polypeptide

<400> 4
Met Lys Trp Val Glu Ser Ile Phe Leu Ile Phe Leu Leu Asn Phe Thr 
1               5                   10                  15      


Glu Ser Arg Thr Leu His Arg Asn Glu Tyr Gly Ile Ala Ser Ile Leu 
            20                  25                  30          


Asp Ser Tyr Gln Cys Thr Ala Glu Ile Ser Leu Ala Asp Leu Ala Thr 
        35                  40                  45              


Ile Phe Phe Ala Gln Phe Val Gln Glu Ala Thr Tyr Lys Glu Val Ser 
    50                  55                  60                  


Lys Met Val Lys Asp Ala Leu Thr Ala Ile Glu Lys Pro Thr Gly Asp 
65                  70                  75                  80  


Glu Gln Ser Ser Gly Cys Leu Glu Asn Gln Leu Pro Ala Phe Leu Glu 
                85                  90                  95      


Glu Leu Cys His Glu Lys Glu Ile Leu Glu Lys Tyr Gly His Ser Asp 
            100                 105                 110         


Cys Cys Ser Gln Ser Glu Glu Gly Arg His Asn Cys Phe Leu Ala His 
        115                 120                 125             


Lys Lys Pro Thr Pro Ala Ser Ile Pro Leu Phe Gln Val Pro Glu Pro 
    130                 135                 140                 


Val Thr Ser Cys Glu Ala Tyr Glu Glu Asp Arg Glu Thr Phe Met Asn 
145                 150                 155                 160 


Lys Phe Ile Tyr Glu Ile Ala Arg Arg His Pro Phe Leu Tyr Ala Pro 
                165                 170                 175     


Thr Ile Leu Leu Trp Ala Ala Arg Tyr Asp Lys Ile Ile Pro Ser Cys 
            180                 185                 190         


Cys Lys Ala Glu Asn Ala Val Glu Cys Phe Gln Thr Lys Ala Ala Thr 
        195                 200                 205             


Val Thr Lys Glu Leu Arg Glu Ser Ser Leu Leu Asn Gln His Ala Cys 
    210                 215                 220                 


Ala Val Met Lys Asn Phe Gly Thr Arg Thr Phe Gln Ala Ile Thr Val 
225                 230                 235                 240 


Thr Lys Leu Ser Gln Lys Phe Thr Lys Val Asn Phe Thr Glu Ile Gln 
                245                 250                 255     


Lys Leu Val Leu Asp Val Ala His Val His Glu His Cys Cys Arg Gly 
            260                 265                 270         


Asp Val Leu Asp Cys Leu Gln Asp Gly Glu Lys Ile Met Ser Tyr Ile 
        275                 280                 285             


Cys Ser Gln Gln Asp Thr Leu Ser Asn Lys Ile Thr Glu Cys Cys Lys 
    290                 295                 300                 


Leu Thr Thr Leu Glu Arg Gly Gln Cys Ile Ile His Ala Glu Asn Asp 
305                 310                 315                 320 


Glu Lys Pro Glu Gly Leu Ser Pro Asn Leu Asn Arg Phe Leu Gly Asp 
                325                 330                 335     


Arg Asp Phe Asn Gln Phe Ser Ser Gly Glu Lys Asn Ile Phe Leu Ala 
            340                 345                 350         


Ser Phe Val His Glu Tyr Ser Arg Arg His Pro Gln Leu Ala Val Ser 
        355                 360                 365             


Val Ile Leu Arg Val Ala Lys Gly Tyr Gln Glu Leu Leu Glu Lys Cys 
    370                 375                 380                 


Phe Gln Thr Glu Asn Pro Leu Glu Cys Gln Asp Lys Gly Glu Glu Glu 
385                 390                 395                 400 


Leu Gln Lys Tyr Ile Gln Glu Ser Gln Ala Leu Ala Lys Arg Ser Cys 
                405                 410                 415     


Gly Leu Phe Gln Lys Leu Gly Glu Tyr Tyr Leu Gln Asn Ala Phe Leu 
            420                 425                 430         


Val Ala Tyr Thr Lys Lys Ala Pro Gln Leu Thr Ser Ser Glu Leu Met 
        435                 440                 445             


Ala Ile Thr Arg Lys Met Ala Ala Thr Ala Ala Thr Cys Cys Gln Leu 
    450                 455                 460                 


Ser Glu Asp Lys Leu Leu Ala Cys Gly Glu Gly Ala Ala Asp Ile Ile 
465                 470                 475                 480 


Ile Gly His Leu Cys Ile Arg His Glu Met Thr Pro Val Asn Pro Gly 
                485                 490                 495     


Val Gly Gln Cys Cys Thr Ser Ser Tyr Ala Asn Arg Arg Pro Cys Phe 
            500                 505                 510         


Ser Ser Leu Val Val Asp Glu Thr Tyr Val Pro Pro Ala Phe Ser Asp 
        515                 520                 525             


Asp Lys Phe Ile Phe His Lys Asp Leu Cys Gln Ala Gln Gly Val Ala 
    530                 535                 540                 


Leu Gln Thr Met Lys Gln Glu Phe Leu Ile Asn Leu Val Lys Gln Lys 
545                 550                 555                 560 


Pro Gln Ile Thr Glu Glu Gln Leu Glu Ala Val Ile Ala Asp Phe 
                565                 570                 575 


<210> 5
<211> 92
<212> PRT
<213> Artificial Sequence

<220>
<223> Description of Artificial Sequence: Synthetic
      polypeptide

<400> 5
Leu Cys Ile Arg His Glu Met Thr Pro Val Asn Pro Gly Val Gly Gln 
1               5                   10                  15      


Cys Cys Thr Ser Ser Tyr Ala Asn Arg Arg Pro Cys Phe Ser Ser Leu 
            20                  25                  30          


Val Val Asp Glu Thr Tyr Val Pro Pro Ala Phe Ser Asp Asp Lys Phe 
        35                  40                  45              


Ile Phe His Lys Asp Leu Cys Gln Ala Gln Gly Val Ala Leu Gln Thr 
    50                  55                  60                  


Met Lys Gln Glu Phe Leu Ile Asn Leu Val Lys Gln Lys Pro Gln Ile 
65                  70                  75                  80  


Thr Glu Glu Gln Leu Glu Ala Val Ile Ala Asp Phe 
                85                  90          


<210> 6
<211> 15
<212> PRT
<213> Homo sapiens

<400> 6
Leu Glu Val Asp Glu Thr Tyr Val Pro Lys Glu Phe Asn Ala Glu 
1               5                   10                  15  


<210> 7
<211> 15
<212> PRT
<213> Homo sapiens

<400> 7
Leu Val Val Asp Glu Thr Tyr Val Pro Pro Ala Phe Ser Asp Asp 
1               5                   10                  15  


<210> 8
<211> 7
<212> PRT
<213> Homo sapiens

<400> 8
Glu Lys Thr Pro Val Ser Asp 
1               5           


<210> 9
<211> 7
<212> PRT
<213> Homo sapiens

<400> 9
Glu Met Thr Pro Val Asn Pro 
1               5           


<210> 10
<211> 54
<212> PRT
<213> Homo sapiens

<400> 10
Pro Gln Val Ser Thr Pro Thr Leu Val Glu Val Ser Arg Asn Leu Gly 
1               5                   10                  15      


Lys Val Gly Ser Lys Cys Cys Lys His Pro Glu Ala Lys Arg Met Pro 
            20                  25                  30          


Cys Ala Glu Asp Tyr Leu Ser Val Val Leu Asn Gln Leu Cys Val Leu 
        35                  40                  45              


His Glu Lys Thr Pro Val 
    50                  


<210> 11
<211> 54
<212> PRT
<213> Homo sapiens

<400> 11
Pro Gln Leu Thr Ser Ser Glu Leu Met Ala Ile Thr Arg Lys Met Ala 
1               5                   10                  15      


Ala Thr Ala Ala Thr Cys Cys Gln Leu Ser Glu Asp Lys Leu Leu Ala 
            20                  25                  30          


Cys Gly Glu Gly Ala Ala Asp Ile Ile Ile Gly His Leu Cys Ile Arg 
        35                  40                  45              


His Glu Met Thr Pro Val 
    50                  


<210> 12
<211> 58
<212> PRT
<213> Homo sapiens

<400> 12
Phe His Ala Asp Ile Cys Thr Leu Ser Glu Lys Glu Arg Gln Ile Lys 
1               5                   10                  15      


Lys Gln Thr Ala Leu Val Glu Leu Val Lys His Lys Pro Lys Ala Thr 
            20                  25                  30          


Lys Glu Gln Leu Lys Ala Val Met Asp Asp Phe Ala Ala Phe Val Glu 
        35                  40                  45              


Lys Cys Cys Lys Ala Asp Asp Lys Glu Thr 
    50                  55              


<210> 13
<211> 58
<212> PRT
<213> Homo sapiens

<400> 13
Phe His Lys Asp Leu Cys Gln Ala Gln Gly Val Ala Leu Gln Thr Met 
1               5                   10                  15      


Lys Gln Glu Phe Leu Ile Asn Leu Val Lys Gln Lys Pro Gln Ile Thr 
            20                  25                  30          


Glu Glu Gln Leu Glu Ala Val Ile Ala Asp Phe Ser Gly Leu Leu Glu 
        35                  40                  45              


Lys Cys Cys Gln Gly Gln Glu Gln Glu Val 
    50                  55              


<210> 14
<211> 31
<212> PRT
<213> Homo sapiens

<400> 14
Gln Leu Cys Val Leu His Glu Lys Thr Pro Val Ser Asp Arg Val Thr 
1               5                   10                  15      


Lys Cys Cys Thr Glu Ser Leu Val Asn Arg Arg Pro Cys Phe Ser 
            20                  25                  30      


<210> 15
<211> 31
<212> PRT
<213> Homo sapiens

<400> 15
His Leu Cys Ile Arg His Glu Met Thr Pro Val Asn Pro Gly Val Gly 
1               5                   10                  15      


Gln Cys Cys Thr Ser Ser Tyr Ala Asn Arg Arg Pro Cys Phe Ser 
            20                  25                  30      


<210> 16
<211> 11
<212> PRT
<213> Homo sapiens

<400> 16
Lys Lys Val Pro Gln Val Ser Thr Pro Thr Leu 
1               5                   10      


<210> 17
<211> 11
<212> PRT
<213> Homo sapiens

<400> 17
Lys Lys Ala Pro Gln Leu Thr Ser Ser Glu Leu 
1               5                   10      


<210> 18
<211> 11
<212> PRT
<213> Homo sapiens

<400> 18
Glu Phe Asn Ala Glu Thr Phe Thr Phe His Ala 
1               5                   10      


<210> 19
<211> 11
<212> PRT
<213> Homo sapiens

<400> 19
Ala Phe Ser Asp Asp Lys Phe Ile Phe His Lys 
1               5                   10      


<210> 20
<211> 11
<212> PRT
<213> Homo sapiens

<400> 20
Pro Lys Glu Phe Asn Ala Glu Thr Phe Thr Phe 
1               5                   10      


<210> 21
<211> 11
<212> PRT
<213> Homo sapiens

<400> 21
Pro Pro Ala Phe Ser Asp Asp Lys Phe Ile Phe 
1               5                   10      


<210> 22
<211> 11
<212> PRT
<213> Homo sapiens

<400> 22
Lys Leu Val Ala Ala Ser Gln Ala Ala Leu Gly 
1               5                   10      


<210> 23
<211> 11
<212> PRT
<213> Homo sapiens

<400> 23
Lys Leu Ile Ser Lys Thr Arg Ala Ala Leu Gly 
1               5                   10      


<210> 24
<211> 9
<212> PRT
<213> Homo sapiens

<400> 24
Cys Phe Ala Glu Glu Gly Lys Lys Leu 
1               5                   


<210> 25
<211> 9
<212> PRT
<213> Homo sapiens

<400> 25
Cys Phe Ala Glu Glu Gly Gln Lys Leu 
1               5                   


<210> 26
<211> 11
<212> PRT
<213> Homo sapiens

<400> 26
Leu Arg Glu Thr Tyr Gly Glu Met Ala Asp Cys 
1               5                   10      


<210> 27
<211> 11
<212> PRT
<213> Homo sapiens

<400> 27
Ile Leu Glu Lys Tyr Gly His Ser Asp Cys Cys 
1               5                   10      


<210> 28
<211> 15
<212> PRT
<213> Homo sapiens

<400> 28
His Lys Asp Asp Asn Pro Asn Leu Pro Arg Leu Val Arg Pro Glu 
1               5                   10                  15  


<210> 29
<211> 15
<212> PRT
<213> Homo sapiens

<400> 29
Lys Lys Pro Thr Pro Ala Ser Ile Pro Leu Phe Gln Val Pro Glu 
1               5                   10                  15  


<210> 30
<211> 49
<212> PRT
<213> Homo sapiens

<400> 30
Thr Phe Thr Phe His Ala Asp Ile Cys Thr Leu Ser Glu Lys Glu Arg 
1               5                   10                  15      


Gln Ile Lys Lys Gln Thr Ala Leu Val Glu Leu Val Lys His Lys Pro 
            20                  25                  30          


Lys Ala Thr Lys Glu Gln Leu Lys Ala Val Met Asp Asp Phe Ala Ala 
        35                  40                  45              


Phe 
    


<210> 31
<211> 49
<212> PRT
<213> Homo sapiens

<400> 31
Lys Phe Ile Phe His Lys Asp Leu Cys Gln Ala Gln Gly Val Ala Leu 
1               5                   10                  15      


Gln Thr Met Lys Gln Glu Phe Leu Ile Asn Leu Val Lys Gln Lys Pro 
            20                  25                  30          


Gln Ile Thr Glu Glu Gln Leu Glu Ala Val Ile Ala Asp Phe Ser Gly 
        35                  40                  45              


Leu 
    


