                         SEQUENCE LISTING

<110>  Autolus Ltd
 
<120>  Antibody conjugate

<130>  P119996PCT

<150>  GB 2004263.6
<151>  2020-03-24

<160>  21    

<170>  PatentIn version 3.5

<210>  1
<211>  124
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  VH domain  (heavy chain variable domain) of humanised JOVI-1 
       antibody, hJOVI-1

<400>  1

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


Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Gly Tyr 
            20                  25                  30          


Val Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 
        35                  40                  45              


Gly Phe Ile Asn Pro Tyr Asn Asp Asp Ile Gln Ser Asn Glu Arg Phe 
    50                  55                  60                  


Arg Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr 
65                  70                  75                  80  


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


Ala Arg Gly Ala Gly Tyr Asn Phe Asp Gly Ala Tyr Arg Phe Phe Asp 
            100                 105                 110         


Phe Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser 
        115                 120                 


<210>  2
<211>  112
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  VL domain  (light chain variable domain) of hJOVI-1

<400>  2

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


Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Arg Leu Val His Ser 
            20                  25                  30          


Asn Gly Asn Thr Tyr Leu His Trp Tyr Leu Gln Lys Pro Gly Gln Ser 
        35                  40                  45              


Pro Arg Leu Leu Ile Tyr Arg Val Ser Asn Arg Phe Pro 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 Ser Gln Ser 
                85                  90                  95      


Thr His Val Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 
            100                 105                 110         


<210>  3
<211>  21
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  signal peptide

<400>  3

Met Gly Thr Ser Leu Leu Cys Trp Met Ala Leu Cys Leu Leu Gly Ala 
1               5                   10                  15      


Asp His Ala Asp Gly 
            20      


<210>  4
<211>  21
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  signal peptide derived from IgG1

<400>  4

Met Ser Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu 
1               5                   10                  15      


His Ala Ala Arg Pro 
            20      


<210>  5
<211>  20
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  signal peptide derived from CD8

<400>  5

Met Ala Val Pro Thr Gln Val Leu Gly Leu Leu Leu Leu Trp Leu Thr 
1               5                   10                  15      


Asp Ala Arg Cys 
            20  


<210>  6
<211>  234
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  spacer sequence, hinge-CH2CH3 of human IgG1

<400>  6

Ala Glu Pro Lys Ser Pro Asp Lys Thr His Thr Cys Pro Pro Cys Pro 
1               5                   10                  15      


Ala Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro 
            20                  25                  30          


Lys Asp Thr Leu Met Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val 
        35                  40                  45              


Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val 
    50                  55                  60                  


Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln 
65                  70                  75                  80  


Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln 
                85                  90                  95      


Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala 
            100                 105                 110         


Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro 
        115                 120                 125             


Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr 
    130                 135                 140                 


Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser 
145                 150                 155                 160 


Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr 
                165                 170                 175     


Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr 
            180                 185                 190         


Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe 
        195                 200                 205             


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


Ser Leu Ser Leu Ser Pro Gly Lys Lys Asp 
225                 230                 


<210>  7
<211>  46
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  spacer sequence, human CD8 stalk

<400>  7

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 
        35                  40                  45      


<210>  8
<211>  20
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  spacer sequence, human IgG1 hinge

<400>  8

Ala Glu Pro Lys Ser Pro Asp Lys Thr His Thr Cys Pro Pro Cys Pro 
1               5                   10                  15      


Lys Asp Pro Lys 
            20  


<210>  9
<211>  185
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  spacer sequence, CD2 ectodomain

<400>  9

Lys Glu Ile Thr Asn Ala Leu Glu Thr Trp Gly Ala Leu Gly Gln Asp 
1               5                   10                  15      


Ile Asn Leu Asp Ile Pro Ser Phe Gln Met Ser Asp Asp Ile Asp Asp 
            20                  25                  30          


Ile Lys Trp Glu Lys Thr Ser Asp Lys Lys Lys Ile Ala Gln Phe Arg 
        35                  40                  45              


Lys Glu Lys Glu Thr Phe Lys Glu Lys Asp Thr Tyr Lys Leu Phe Lys 
    50                  55                  60                  


Asn Gly Thr Leu Lys Ile Lys His Leu Lys Thr Asp Asp Gln Asp Ile 
65                  70                  75                  80  


Tyr Lys Val Ser Ile Tyr Asp Thr Lys Gly Lys Asn Val Leu Glu Lys 
                85                  90                  95      


Ile Phe Asp Leu Lys Ile Gln Glu Arg Val Ser Lys Pro Lys Ile Ser 
            100                 105                 110         


Trp Thr Cys Ile Asn Thr Thr Leu Thr Cys Glu Val Met Asn Gly Thr 
        115                 120                 125             


Asp Pro Glu Leu Asn Leu Tyr Gln Asp Gly Lys His Leu Lys Leu Ser 
    130                 135                 140                 


Gln Arg Val Ile Thr His Lys Trp Thr Thr Ser Leu Ser Ala Lys Phe 
145                 150                 155                 160 


Lys Cys Thr Ala Gly Asn Lys Val Ser Lys Glu Ser Ser Val Glu Pro 
                165                 170                 175     


Val Ser Cys Pro Glu Lys Gly Leu Asp 
            180                 185 


<210>  10
<211>  259
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  spacer sequence, CD34 ectodomain

<400>  10

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


Thr Phe Ser Asn Val Ser Thr Asn Val Ser Tyr Gln Glu Thr Thr Thr 
            20                  25                  30          


Pro Ser Thr Leu Gly Ser Thr Ser Leu His Pro Val Ser Gln His Gly 
        35                  40                  45              


Asn Glu Ala Thr Thr Asn Ile Thr Glu Thr Thr Val Lys Phe Thr Ser 
    50                  55                  60                  


Thr Ser Val Ile Thr Ser Val Tyr Gly Asn Thr Asn Ser Ser Val Gln 
65                  70                  75                  80  


Ser Gln Thr Ser Val Ile Ser Thr Val Phe Thr Thr Pro Ala Asn Val 
                85                  90                  95      


Ser Thr Pro Glu Thr Thr Leu Lys Pro Ser Leu Ser Pro Gly Asn Val 
            100                 105                 110         


Ser Asp Leu Ser Thr Thr Ser Thr Ser Leu Ala Thr Ser Pro Thr Lys 
        115                 120                 125             


Pro Tyr Thr Ser Ser Ser Pro Ile Leu Ser Asp Ile Lys Ala Glu Ile 
    130                 135                 140                 


Lys Cys Ser Gly Ile Arg Glu Val Lys Leu Thr Gln Gly Ile Cys Leu 
145                 150                 155                 160 


Glu Gln Asn Lys Thr Ser Ser Cys Ala Glu Phe Lys Lys Asp Arg Gly 
                165                 170                 175     


Glu Gly Leu Ala Arg Val Leu Cys Gly Glu Glu Gln Ala Asp Ala Asp 
            180                 185                 190         


Ala Gly Ala Gln Val Cys Ser Leu Leu Leu Ala Gln Ser Glu Val Arg 
        195                 200                 205             


Pro Gln Cys Leu Leu Leu Val Leu Ala Asn Arg Thr Glu Ile Ser Ser 
    210                 215                 220                 


Lys Leu Gln Leu Met Lys Lys His Gln Ser Asp Leu Lys Lys Leu Gly 
225                 230                 235                 240 


Ile Leu Asp Phe Thr Glu Gln Asp Val Ala Ser His Gln Ser Tyr Ser 
                245                 250                 255     


Gln Lys Thr 
            


<210>  11
<211>  21
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  CD8a transmembrane domain

<400>  11

Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu 
1               5                   10                  15      


Ser Leu Val Ile Thr 
            20      


<210>  12
<211>  24
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  TYRP-1 transmembrane domain

<400>  12

Ile Ile Ala Ile Ala Val Val Gly Ala Leu Leu Leu Val Ala Leu Ile 
1               5                   10                  15      


Phe Gly Thr Ala Ser Tyr Leu Ile 
            20                  


<210>  13
<211>  140
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  CD28 transmembrane domain and CD3 Zeta endodomain

<400>  13

Phe Trp Val Leu Val Val Val Gly Gly Val Leu Ala Cys Tyr Ser Leu 
1               5                   10                  15      


Leu Val Thr Val Ala Phe Ile Ile Phe Trp Val Arg Arg Val Lys Phe 
            20                  25                  30          


Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu 
        35                  40                  45              


Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp 
    50                  55                  60                  


Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys 
65                  70                  75                  80  


Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala 
                85                  90                  95      


Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys 
            100                 105                 110         


Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr 
        115                 120                 125             


Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg 
    130                 135                 140 


<210>  14
<211>  180
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  CD28 transmembrane domain and CD28 and CD3 Zeta endodomains

<400>  14

Phe Trp Val Leu Val Val Val Gly Gly Val Leu Ala Cys Tyr Ser Leu 
1               5                   10                  15      


Leu Val Thr Val Ala Phe Ile Ile Phe Trp Val Arg Ser Lys Arg Ser 
            20                  25                  30          


Arg Leu Leu His Ser Asp Tyr Met Asn Met Thr Pro Arg Arg Pro Gly 
        35                  40                  45              


Pro Thr Arg Lys His Tyr Gln Pro Tyr Ala Pro Pro Arg Asp Phe Ala 
    50                  55                  60                  


Ala Tyr Arg Ser Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala 
65                  70                  75                  80  


Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg 
                85                  90                  95      


Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu 
            100                 105                 110         


Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn 
        115                 120                 125             


Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met 
    130                 135                 140                 


Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly 
145                 150                 155                 160 


Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala 
                165                 170                 175     


Leu Pro Pro Arg 
            180 


<210>  15
<211>  216
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  CD28 transmembrane domain and CD28, OX40 and CD3 Zeta endodomains

<400>  15

Phe Trp Val Leu Val Val Val Gly Gly Val Leu Ala Cys Tyr Ser Leu 
1               5                   10                  15      


Leu Val Thr Val Ala Phe Ile Ile Phe Trp Val Arg Ser Lys Arg Ser 
            20                  25                  30          


Arg Leu Leu His Ser Asp Tyr Met Asn Met Thr Pro Arg Arg Pro Gly 
        35                  40                  45              


Pro Thr Arg Lys His Tyr Gln Pro Tyr Ala Pro Pro Arg Asp Phe Ala 
    50                  55                  60                  


Ala Tyr Arg Ser Arg Asp Gln Arg Leu Pro Pro Asp Ala His Lys Pro 
65                  70                  75                  80  


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


Ala His Ser Thr Leu Ala Lys Ile Arg Val Lys Phe Ser Arg Ser Ala 
            100                 105                 110         


Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu 
        115                 120                 125             


Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly 
    130                 135                 140                 


Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu 
145                 150                 155                 160 


Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser 
                165                 170                 175     


Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly 
            180                 185                 190         


Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu 
        195                 200                 205             


His Met Gln Ala Leu Pro Pro Arg 
    210                 215     


<210>  16
<211>  136
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  CD8a transmembrane domain and CD3 Zeta endodomain

<400>  16

Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu 
1               5                   10                  15      


Ser Leu Val Ile Thr Arg Val Leu Tyr Cys Lys Phe Ser Arg Ser Ala 
            20                  25                  30          


Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu 
        35                  40                  45              


Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly 
    50                  55                  60                  


Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu 
65                  70                  75                  80  


Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser 
                85                  90                  95      


Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly 
            100                 105                 110         


Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu 
        115                 120                 125             


His Met Gln Ala Leu Pro Pro Arg 
    130                 135     


<210>  17
<211>  178
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  CD8a transmembrane domain and 4-1BB and CD3 Zeta endodomains

<400>  17

Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu 
1               5                   10                  15      


Ser Leu Val Ile Thr Leu Tyr Cys Lys Arg Gly Arg Lys Lys Leu Leu 
            20                  25                  30          


Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro Val Gln Thr Thr Gln Glu 
        35                  40                  45              


Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu Glu Glu Glu Gly Gly Cys 
    50                  55                  60                  


Glu Leu Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln 
65                  70                  75                  80  


Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu 
                85                  90                  95      


Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly 
            100                 105                 110         


Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu 
        115                 120                 125             


Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly 
    130                 135                 140                 


Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser 
145                 150                 155                 160 


Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro 
                165                 170                 175     


Pro Arg 
        


<210>  18
<211>  178
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  TYRP-1 transmembrane domain and 4-1BB and CD3 Zeta endodomains

<400>  18

Ile Ile Ala Ile Ala Val Val Gly Ala Leu Leu Leu Val Ala Leu Ile 
1               5                   10                  15      


Phe Gly Thr Ala Ser Tyr Leu Ile Lys Arg Gly Arg Lys Lys Leu Leu 
            20                  25                  30          


Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro Val Gln Thr Thr Gln Glu 
        35                  40                  45              


Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu Glu Glu Glu Gly Gly Cys 
    50                  55                  60                  


Glu Leu Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln 
65                  70                  75                  80  


Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu 
                85                  90                  95      


Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly 
            100                 105                 110         


Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu 
        115                 120                 125             


Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly 
    130                 135                 140                 


Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser 
145                 150                 155                 160 


Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro 
                165                 170                 175     


Pro Arg 
        


<210>  19
<211>  45
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  spacer sequence, coiled-coil COMP

<400>  19

Asp Leu Gly Pro Gln Met Leu Arg Glu Leu Gln Glu Thr Asn Ala Ala 
1               5                   10                  15      


Leu Gln Asp Val Arg Glu Leu Leu Arg Gln Gln Val Arg Glu Ile Thr 
            20                  25                  30          


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


<210>  20
<211>  5
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  spacer sequence, truncated coiled-coil COMP

<400>  20

Cys Asp Ala Cys Gly 
1               5   


<210>  21
<211>  20
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  spacer sequence, truncated coiled-coil COMP (C-terminus)

<400>  21

Gln Gln Val Arg Glu Ile Thr Phe Leu Lys Asn Thr Val Met Glu Cys 
1               5                   10                  15      


Asp Ala Cys Gly 
            20  


