                         SEQUENCE LISTING

<110>  Helsingin Yliopisto
 
<120>  A cross-hybrid Fc-fusion polypeptide targeting PD-L1 and methods 
       and uses related thereto

<130>  BP302463

<160>  9     

<170>  PatentIn version 3.5

<210>  1
<211>  385
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Fc-fusion polypeptide

<400>  1

Met Glu Phe Gly Leu Ser Trp Val Phe Leu Val Ala Leu Phe Arg Gly 
1               5                   10                  15      


Val Gln Cys Asp Ser Pro Asp Arg Pro Trp Asn Pro Pro Thr Phe Ser 
            20                  25                  30          


Pro Ala Leu Leu Val Val Thr Glu Gly Asp Asn Ala Thr Phe Thr Cys 
        35                  40                  45              


Ser Phe Ser Asn Thr Ser Glu Ser Phe His Val Val Trp His Arg Glu 
    50                  55                  60                  


Ser Pro Ser Gly Gln Thr Asp Thr Leu Ala Ala Phe Pro Glu Asp Arg 
65                  70                  75                  80  


Ser Gln Pro Gly Gln Asp Cys Arg Phe Arg Val Thr Gln Leu Pro Asn 
                85                  90                  95      


Gly Arg Asp Phe His Met Ser Val Val Arg Ala Arg Arg Asn Asp Ser 
            100                 105                 110         


Gly Thr Tyr Val Cys Gly Val Ile Ser Leu Ala Pro Lys Ala Gln Ile 
        115                 120                 125             


Lys Glu Ser Leu Arg Ala Glu Leu Arg Val Thr Glu Arg Gly Gly Gly 
    130                 135                 140                 


Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Pro Ala Pro Glu 
145                 150                 155                 160 


Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Ala Leu Glu Asp 
                165                 170                 175     


Leu Leu Leu Gly Ser Glu Ala Asn Gly Val Thr Cys Val Val Val Asp 
            180                 185                 190         


Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 
        195                 200                 205             


Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 
    210                 215                 220                 


Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 
225                 230                 235                 240 


Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 
                245                 250                 255     


Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Ser Gly Asn Thr Phe 
            260                 265                 270         


Arg Pro Glu Val His Leu Leu Pro Pro Pro Ser Glu Glu Leu Ala Leu 
        275                 280                 285             


Asn Glu Leu Val Thr Leu Thr Cys Leu Ala Arg Gly Phe Ser Pro Lys 
    290                 295                 300                 


Asp Val Leu Val Arg Trp Leu Gln Gly Ser Gln Glu Leu Pro Arg Glu 
305                 310                 315                 320 


Lys Tyr Leu Thr Trp Ala Ser Arg Gln Glu Pro Ser Gln Gly Thr Thr 
                325                 330                 335     


Thr Phe Ala Val Thr Ser Ile Leu Arg Val Ala Ala Glu Asp Trp Lys 
            340                 345                 350         


Lys Gly Asp Thr Phe Ser Cys Met Val Gly His Glu Ala Leu Pro Leu 
        355                 360                 365             


Ala Phe Thr Gln Lys Thr Ile Asp Arg His His His His His His His 
    370                 375                 380                 


His 
385 


<210>  2
<211>  122
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  PD-1 ectodomain

<400>  2

Asp Ser Pro Asp Arg Pro Trp Asn Pro Pro Thr Phe Ser Pro Ala Leu 
1               5                   10                  15      


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


Asn Thr Ser Glu Ser Phe His Val Val Trp His Arg Glu Ser Pro Ser 
        35                  40                  45              


Gly Gln Thr Asp Thr Leu Ala Ala Phe Pro Glu Asp Arg Ser Gln Pro 
    50                  55                  60                  


Gly Gln Asp Cys Arg Phe Arg Val Thr Gln Leu Pro Asn Gly Arg Asp 
65                  70                  75                  80  


Phe His Met Ser Val Val Arg Ala Arg Arg Asn Asp Ser Gly Thr Tyr 
                85                  90                  95      


Val Cys Gly Val Ile Ser Leu Ala Pro Lys Ala Gln Ile Lys Glu Ser 
            100                 105                 110         


Leu Arg Ala Glu Leu Arg Val Thr Glu Arg 
        115                 120         


<210>  3
<211>  221
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Fc of IgGA

<400>  3

Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro 
1               5                   10                  15      


Ala Leu Glu Asp Leu Leu Leu Gly Ser Glu Ala Asn Gly Val Thr Cys 
            20                  25                  30          


Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 
        35                  40                  45              


Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 
    50                  55                  60                  


Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 
65                  70                  75                  80  


His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 
                85                  90                  95      


Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Ser 
            100                 105                 110         


Gly Asn Thr Phe Arg Pro Glu Val His Leu Leu Pro Pro Pro Ser Glu 
        115                 120                 125             


Glu Leu Ala Leu Asn Glu Leu Val Thr Leu Thr Cys Leu Ala Arg Gly 
    130                 135                 140                 


Phe Ser Pro Lys Asp Val Leu Val Arg Trp Leu Gln Gly Ser Gln Glu 
145                 150                 155                 160 


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


Gln Gly Thr Thr Thr Phe Ala Val Thr Ser Ile Leu Arg Val Ala Ala 
            180                 185                 190         


Glu Asp Trp Lys Lys Gly Asp Thr Phe Ser Cys Met Val Gly His Glu 
        195                 200                 205             


Ala Leu Pro Leu Ala Phe Thr Gln Lys Thr Ile Asp Arg 
    210                 215                 220     


<210>  4
<211>  385
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Modified (DF+TE) Fc-fusion polypeptide

<400>  4

Met Glu Phe Gly Leu Ser Trp Val Phe Leu Val Ala Leu Phe Arg Gly 
1               5                   10                  15      


Val Gln Cys Asp Ser Pro Asp Arg Pro Trp Asn Pro Pro Thr Phe Ser 
            20                  25                  30          


Pro Ala Leu Leu Val Val Thr Glu Gly Asp Asn Ala Thr Phe Thr Cys 
        35                  40                  45              


Ser Phe Ser Asn Thr Ser Glu Ser Phe His Val Val Trp His Arg Glu 
    50                  55                  60                  


Ser Pro Ser Gly Gln Thr Asp Thr Leu Ala Ala Phe Pro Glu Asp Arg 
65                  70                  75                  80  


Ser Gln Pro Gly Gln Asp Cys Arg Phe Arg Val Thr Gln Leu Pro Asn 
                85                  90                  95      


Gly Arg Asp Phe His Met Ser Val Val Arg Ala Arg Arg Asn Asp Ser 
            100                 105                 110         


Gly Thr Tyr Val Cys Gly Val Ile Ser Leu Ala Pro Lys Ala Gln Ile 
        115                 120                 125             


Lys Glu Ser Leu Arg Ala Glu Leu Arg Val Thr Glu Arg Gly Gly Gly 
    130                 135                 140                 


Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Pro Ala Pro Glu 
145                 150                 155                 160 


Leu Leu Gly Gly Pro Asp Val Phe Leu Phe Pro Pro Ala Leu Glu Asp 
                165                 170                 175     


Leu Leu Leu Gly Ser Glu Ala Asn Gly Val Thr Cys Val Val Val Asp 
            180                 185                 190         


Val Ser Phe Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 
        195                 200                 205             


Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 
    210                 215                 220                 


Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 
225                 230                 235                 240 


Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Thr Asn Lys Ala Leu Pro 
                245                 250                 255     


Ala Pro Glu Glu Lys Thr Ile Ser Lys Ala Lys Ser Gly Asn Thr Phe 
            260                 265                 270         


Arg Pro Glu Val His Leu Leu Pro Pro Pro Ser Glu Glu Leu Ala Leu 
        275                 280                 285             


Asn Glu Leu Val Thr Leu Thr Cys Leu Ala Arg Gly Phe Ser Pro Lys 
    290                 295                 300                 


Asp Val Leu Val Arg Trp Leu Gln Gly Ser Gln Glu Leu Pro Arg Glu 
305                 310                 315                 320 


Lys Tyr Leu Thr Trp Ala Ser Arg Gln Glu Pro Ser Gln Gly Thr Thr 
                325                 330                 335     


Thr Phe Ala Val Thr Ser Ile Leu Arg Val Ala Ala Glu Asp Trp Lys 
            340                 345                 350         


Lys Gly Asp Thr Phe Ser Cys Met Val Gly His Glu Ala Leu Pro Leu 
        355                 360                 365             


Ala Phe Thr Gln Lys Thr Ile Asp Arg His His His His His His His 
    370                 375                 380                 


His 
385 


<210>  5
<211>  221
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Modified (DF+TE) Fc of IgGA

<400>  5

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


Ala Leu Glu Asp Leu Leu Leu Gly Ser Glu Ala Asn Gly Val Thr Cys 
            20                  25                  30          


Val Val Val Asp Val Ser Phe Glu Asp Pro Glu Val Lys Phe Asn Trp 
        35                  40                  45              


Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 
    50                  55                  60                  


Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 
65                  70                  75                  80  


His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Thr Asn 
                85                  90                  95      


Lys Ala Leu Pro Ala Pro Glu Glu Lys Thr Ile Ser Lys Ala Lys Ser 
            100                 105                 110         


Gly Asn Thr Phe Arg Pro Glu Val His Leu Leu Pro Pro Pro Ser Glu 
        115                 120                 125             


Glu Leu Ala Leu Asn Glu Leu Val Thr Leu Thr Cys Leu Ala Arg Gly 
    130                 135                 140                 


Phe Ser Pro Lys Asp Val Leu Val Arg Trp Leu Gln Gly Ser Gln Glu 
145                 150                 155                 160 


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


Gln Gly Thr Thr Thr Phe Ala Val Thr Ser Ile Leu Arg Val Ala Ala 
            180                 185                 190         


Glu Asp Trp Lys Lys Gly Asp Thr Phe Ser Cys Met Val Gly His Glu 
        195                 200                 205             


Ala Leu Pro Leu Ala Phe Thr Gln Lys Thr Ile Asp Arg 
    210                 215                 220     


<210>  6
<211>  663
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Polynucleotide encoding the Fc region of the fusion polypeptide

<400>  6
cctgctccag aactgctcgg cggaccttcc gtgttcctgt ttccaccagc tctggaagat       60

ctgctgctgg gctctgaagc caatggcgtg acatgtgtgg tggtggacgt gtcccacgag      120

gaccccgaag tgaagttcaa ttggtacgtg gacggcgtgg aagtgcacaa cgccaagacc      180

aagcctagag aggaacagta caacagcacc tacagagtgg tgtccgtgct gaccgtgctg      240

caccaggatt ggctgaacgg caaagagtac aagtgcaagg tgtccaacaa ggccctgcct      300

gctcctatcg agaaaaccat cagcaaggcc aagagcggca acaccttcag acccgaagtg      360

catctgctgc ctccacctag cgaagaactg gccctgaatg agctggtcac cctgacatgt      420

ctggccaggg gcttcagccc taaggatgtg ctcgttagat ggctgcaggg cagccaagag      480

ctgcccagag agaagtatct gacctgggcc agcagacaag agcctagcca gggcaccaca      540

acctttgccg tgaccagcat tctgagagtg gccgccgagg attggaagaa gggcgatacc      600

ttcagctgca tggtcggaca cgaagccctg cctctggcct tcacacagaa aacaatcgac      660

cgg                                                                    663


<210>  7
<211>  366
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Polynucleotide encoding the region of PD-1 of the fusion 
       polypeptide

<400>  7
gacagccctg acagaccttg gaaccctcca acattcagcc ccgctctgct ggtggttacc       60

gagggcgata atgccacctt cacctgtagc ttcagcaaca ccagcgagag cttccacgtc      120

gtgtggcaca gagagtctcc aagcggccag acagataccc tggctgcctt tcctgaggac      180

agatctcagc ccggccagga ctgcagattc agagtgaccc agctgcctaa cggccgggac      240

tttcacatga gcgtcgtgcg ggccagaaga aacgatagcg gcacatacgt gtgcggcgtg      300

atctctctgg cccctaaggc tcagatcaaa gagagcctga gagccgagct gagagtgacc      360

gaaaga                                                                 366


<210>  8
<211>  1158
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Polynucleotide encoding the Fc-fusion polypeptide

<400>  8
atggaatttg gcctgagctg ggtgttcctg gtggccctgt ttagaggcgt gcagtgcgac       60

agccctgaca gaccttggaa ccctccaaca ttcagccccg ctctgctggt ggttaccgag      120

ggcgataatg ccaccttcac ctgtagcttc agcaacacca gcgagagctt ccacgtcgtg      180

tggcacagag agtctccaag cggccagaca gataccctgg ctgcctttcc tgaggacaga      240

tctcagcccg gccaggactg cagattcaga gtgacccagc tgcctaacgg ccgggacttt      300

cacatgagcg tcgtgcgggc cagaagaaac gatagcggca catacgtgtg cggcgtgatc      360

tctctggccc ctaaggctca gatcaaagag agcctgagag ccgagctgag agtgaccgaa      420

agaggtggcg gaggatctgg cggaggtgga agcggcggag gcggatctga taagacccac      480

acctgtcctc catgtcctgc tccagaactg ctcggcggac cttccgtgtt cctgtttcca      540

ccagctctgg aagatctgct gctgggctct gaagccaatg gcgtgacatg tgtggtggtg      600

gacgtgtccc acgaggaccc cgaagtgaag ttcaattggt acgtggacgg cgtggaagtg      660

cacaacgcca agaccaagcc tagagaggaa cagtacaaca gcacctacag agtggtgtcc      720

gtgctgaccg tgctgcacca ggattggctg aacggcaaag agtacaagtg caaggtgtcc      780

aacaaggccc tgcctgctcc tatcgagaaa accatcagca aggccaagag cggcaacacc      840

ttcagacccg aagtgcatct gctgcctcca cctagcgaag aactggccct gaatgagctg      900

gtcaccctga catgtctggc caggggcttc agccctaagg atgtgctcgt tagatggctg      960

cagggcagcc aagagctgcc cagagagaag tatctgacct gggccagcag acaagagcct     1020

agccagggca ccacaacctt tgccgtgacc agcattctga gagtggccgc cgaggattgg     1080

aagaagggcg ataccttcag ctgcatggtc ggacacgaag ccctgcctct ggccttcaca     1140

cagaaaacaa tcgaccgg                                                   1158


<210>  9
<211>  1158
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Polynucleotide encoding the modified (DF+TE) Fc-fusion 
       polypeptide

<400>  9
atggaatttg gcctgagctg ggtgttcctg gtggccctgt ttagaggcgt gcagtgcgac       60

agccctgaca gaccttggaa ccctccaaca ttcagccccg ctctgctggt ggttaccgag      120

ggcgataatg ccaccttcac ctgtagcttc agcaacacca gcgagagctt ccacgtcgtg      180

tggcacagag agtctccaag cggccagaca gataccctgg ctgcctttcc tgaggacaga      240

tctcagcccg gccaggactg cagattcaga gtgacccagc tgcctaacgg ccgggacttt      300

cacatgagcg tcgtgcgggc cagaagaaac gatagcggca catacgtgtg cggcgtgatc      360

tctctggccc ctaaggctca gatcaaagag agcctgagag ccgagctgag agtgaccgaa      420

agaggtggcg gaggatctgg cggaggtgga agcggcggag gcggatctga taagacccac      480

acctgtcctc catgtcctgc tccagaactg ctcggcggac ccgatgtgtt cctgtttcca      540

ccagctctgg aagatctgct gctgggctct gaagccaatg gcgtgacatg tgtggtggtg      600

gacgtgtcct tcgaggaccc cgaagtgaag ttcaattggt acgtggacgg cgtggaagtg      660

cacaacgcca agaccaagcc tagagaggaa cagtacaaca gcacctacag agtggtgtcc      720

gtgctgaccg tgctgcacca ggattggctg aacggcaaag agtacaagtg caaagtgacc      780

aacaaggccc tgcctgctcc tgaggaaaag accatcagca aggccaagag cggcaacacc      840

ttcagacccg aagtgcatct gctgcctcca cctagcgaag aactggccct gaatgagctg      900

gtcaccctga catgtctggc caggggcttc agccctaagg atgtgctcgt tagatggctg      960

cagggcagcc aagagctgcc cagagagaag tatctgacct gggccagcag acaagagcct     1020

agccagggca ccacaacctt tgccgtgacc agcattctga gagtggccgc cgaggattgg     1080

aagaagggcg ataccttcag ctgcatggtc ggacacgaag ccctgcctct ggccttcaca     1140

cagaaaacca tcgaccgg                                                   1158


