                         SEQUENCE LISTING

<110>  F. Hoffmann-La Roche AG
 
<120>  On-column enzymatic cleavage

<130>  30890 WO

<150>  EP 12157510.4
<151>  2012-02-29

<160>  21    

<170>  PatentIn version 3.5

<210>  1
<211>  309
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  interferon and tag and cleavage site containing 
       tetranectin-apolipoprotein A-I fusion polypeptide

<400>  1

Cys Asp Leu Pro Gln Thr His Ser Leu Gly Ser His His His His His 
1               5                   10                  15      


His Gly Ser Val Val Ala Pro Pro Ala Pro Ile Val Asn Ala Lys Lys 
            20                  25                  30          


Asp Val Val Asn Thr Lys Met Phe Glu Glu Leu Lys Ser Arg Leu Asp 
        35                  40                  45              


Thr Leu Ala Gln Glu Val Ala Leu Leu Lys Glu Gln Gln Ala Leu Gln 
    50                  55                  60                  


Thr Val Asp Glu Pro Pro Gln Ser Pro Trp Asp Arg Val Lys Asp Leu 
65                  70                  75                  80  


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


Ser Gln Phe Glu Gly Ser Ala Leu Gly Lys Gln Leu Asn Leu Lys Leu 
            100                 105                 110         


Leu Asp Asn Trp Asp Ser Val Thr Ser Thr Phe Ser Lys Leu Arg Glu 
        115                 120                 125             


Gln Leu Gly Pro Val Thr Gln Glu Phe Trp Asp Asn Leu Glu Lys Glu 
    130                 135                 140                 


Thr Glu Gly Leu Arg Gln Glu Met Ser Lys Asp Leu Glu Glu Val Lys 
145                 150                 155                 160 


Ala Lys Val Gln Pro Tyr Leu Asp Asp Phe Gln Lys Lys Trp Gln Glu 
                165                 170                 175     


Glu Met Glu Leu Tyr Arg Gln Lys Val Glu Pro Leu Arg Ala Glu Leu 
            180                 185                 190         


Gln Glu Gly Ala Arg Gln Lys Leu His Glu Leu Gln Glu Lys Leu Ser 
        195                 200                 205             


Pro Leu Gly Glu Glu Met Arg Asp Arg Ala Arg Ala His Val Asp Ala 
    210                 215                 220                 


Leu Arg Thr His Leu Ala Pro Tyr Ser Asp Glu Leu Arg Gln Arg Leu 
225                 230                 235                 240 


Ala Ala Arg Leu Glu Ala Leu Lys Glu Asn Gly Gly Ala Arg Leu Ala 
                245                 250                 255     


Glu Tyr His Ala Lys Ala Thr Glu His Leu Ser Thr Leu Ser Glu Lys 
            260                 265                 270         


Ala Lys Pro Ala Leu Glu Asp Leu Arg Gln Gly Leu Leu Pro Val Leu 
        275                 280                 285             


Glu Ser Phe Lys Val Ser Phe Leu Ser Ala Leu Glu Glu Tyr Thr Lys 
    290                 295                 300                 


Lys Leu Asn Thr Gln 
305                 


<210>  2
<211>  302
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  short leader and tag and cleavage site containing 
       tetranectin-apolipoprotein A-I fusion polypeptide

<400>  2

Met Arg Gly Ser His His His His His His Gly Ser Val Val Ala Pro 
1               5                   10                  15      


Pro Ala Pro Ile Val Asn Ala Lys Lys Asp Val Val Asn Thr Lys Met 
            20                  25                  30          


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


Leu Leu Lys Glu Gln Gln Ala Leu Gln Thr Val Asp Glu Pro Pro Gln 
    50                  55                  60                  


Ser Pro Trp Asp Arg Val Lys Asp Leu Ala Thr Val Tyr Val Asp Val 
65                  70                  75                  80  


Leu Lys Asp Ser Gly Arg Asp Tyr Val Ser Gln Phe Glu Gly Ser Ala 
                85                  90                  95      


Leu Gly Lys Gln Leu Asn Leu Lys Leu Leu Asp Asn Trp Asp Ser Val 
            100                 105                 110         


Thr Ser Thr Phe Ser Lys Leu Arg Glu Gln Leu Gly Pro Val Thr Gln 
        115                 120                 125             


Glu Phe Trp Asp Asn Leu Glu Lys Glu Thr Glu Gly Leu Arg Gln Glu 
    130                 135                 140                 


Met Ser Lys Asp Leu Glu Glu Val Lys Ala Lys Val Gln Pro Tyr Leu 
145                 150                 155                 160 


Asp Asp Phe Gln Lys Lys Trp Gln Glu Glu Met Glu Leu Tyr Arg Gln 
                165                 170                 175     


Lys Val Glu Pro Leu Arg Ala Glu Leu Gln Glu Gly Ala Arg Gln Lys 
            180                 185                 190         


Leu His Glu Leu Gln Glu Lys Leu Ser Pro Leu Gly Glu Glu Met Arg 
        195                 200                 205             


Asp Arg Ala Arg Ala His Val Asp Ala Leu Arg Thr His Leu Ala Pro 
    210                 215                 220                 


Tyr Ser Asp Glu Leu Arg Gln Arg Leu Ala Ala Arg Leu Glu Ala Leu 
225                 230                 235                 240 


Lys Glu Asn Gly Gly Ala Arg Leu Ala Glu Tyr His Ala Lys Ala Thr 
                245                 250                 255     


Glu His Leu Ser Thr Leu Ser Glu Lys Ala Lys Pro Ala Leu Glu Asp 
            260                 265                 270         


Leu Arg Gln Gly Leu Leu Pro Val Leu Glu Ser Phe Lys Val Ser Phe 
        275                 280                 285             


Leu Ser Ala Leu Glu Glu Tyr Thr Lys Lys Leu Asn Thr Gln 
    290                 295                 300         


<210>  3
<211>  298
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  tag and cleavage site containing tetranectin-apolipoprotein A-I 
       fusion polypeptide

<400>  3

His His His His His His Gly Ser Val Val Ala Pro Pro Ala Pro Ile 
1               5                   10                  15      


Val Asn Ala Lys Lys Asp Val Val Asn Thr Lys Met Phe Glu Glu Leu 
            20                  25                  30          


Lys Ser Arg Leu Asp Thr Leu Ala Gln Glu Val Ala Leu Leu Lys Glu 
        35                  40                  45              


Gln Gln Ala Leu Gln Thr Val Asp Glu Pro Pro Gln Ser Pro Trp Asp 
    50                  55                  60                  


Arg Val Lys Asp Leu Ala Thr Val Tyr Val Asp Val Leu Lys Asp Ser 
65                  70                  75                  80  


Gly Arg Asp Tyr Val Ser Gln Phe Glu Gly Ser Ala Leu Gly Lys Gln 
                85                  90                  95      


Leu Asn Leu Lys Leu Leu Asp Asn Trp Asp Ser Val Thr Ser Thr Phe 
            100                 105                 110         


Ser Lys Leu Arg Glu Gln Leu Gly Pro Val Thr Gln Glu Phe Trp Asp 
        115                 120                 125             


Asn Leu Glu Lys Glu Thr Glu Gly Leu Arg Gln Glu Met Ser Lys Asp 
    130                 135                 140                 


Leu Glu Glu Val Lys Ala Lys Val Gln Pro Tyr Leu Asp Asp Phe Gln 
145                 150                 155                 160 


Lys Lys Trp Gln Glu Glu Met Glu Leu Tyr Arg Gln Lys Val Glu Pro 
                165                 170                 175     


Leu Arg Ala Glu Leu Gln Glu Gly Ala Arg Gln Lys Leu His Glu Leu 
            180                 185                 190         


Gln Glu Lys Leu Ser Pro Leu Gly Glu Glu Met Arg Asp Arg Ala Arg 
        195                 200                 205             


Ala His Val Asp Ala Leu Arg Thr His Leu Ala Pro Tyr Ser Asp Glu 
    210                 215                 220                 


Leu Arg Gln Arg Leu Ala Ala Arg Leu Glu Ala Leu Lys Glu Asn Gly 
225                 230                 235                 240 


Gly Ala Arg Leu Ala Glu Tyr His Ala Lys Ala Thr Glu His Leu Ser 
                245                 250                 255     


Thr Leu Ser Glu Lys Ala Lys Pro Ala Leu Glu Asp Leu Arg Gln Gly 
            260                 265                 270         


Leu Leu Pro Val Leu Glu Ser Phe Lys Val Ser Phe Leu Ser Ala Leu 
        275                 280                 285             


Glu Glu Tyr Thr Lys Lys Leu Asn Thr Gln 
    290                 295             


<210>  4
<211>  283
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  tetranectin-apolipoprotein A-I fusion polypeptide


<220>
<221>  MISC_FEATURE
<222>  (1)..(1)
<223>  X = A or G or S or T

<400>  4

Ile Val Asn Ala Lys Lys Asp Val Val Asn Thr Lys Met Phe Glu Glu 
1               5                   10                  15      


Leu Lys Ser Arg Leu Asp Thr Leu Ala Gln Glu Val Ala Leu Leu Lys 
            20                  25                  30          


Glu Gln Gln Ala Leu Gln Thr Val Asp Glu Pro Pro Gln Ser Pro Trp 
        35                  40                  45              


Asp Arg Val Lys Asp Leu Ala Thr Val Tyr Val Asp Val Leu Lys Asp 
    50                  55                  60                  


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


Gln Leu Asn Leu Lys Leu Leu Asp Asn Trp Asp Ser Val Thr Ser Thr 
                85                  90                  95      


Phe Ser Lys Leu Arg Glu Gln Leu Gly Pro Val Thr Gln Glu Phe Trp 
            100                 105                 110         


Asp Asn Leu Glu Lys Glu Thr Glu Gly Leu Arg Gln Glu Met Ser Lys 
        115                 120                 125             


Asp Leu Glu Glu Val Lys Ala Lys Val Gln Pro Tyr Leu Asp Asp Phe 
    130                 135                 140                 


Gln Lys Lys Trp Gln Glu Glu Met Glu Leu Tyr Arg Gln Lys Val Glu 
145                 150                 155                 160 


Pro Leu Arg Ala Glu Leu Gln Glu Gly Ala Arg Gln Lys Leu His Glu 
                165                 170                 175     


Leu Gln Glu Lys Leu Ser Pro Leu Gly Glu Glu Met Arg Asp Arg Ala 
            180                 185                 190         


Arg Ala His Val Asp Ala Leu Arg Thr His Leu Ala Pro Tyr Ser Asp 
        195                 200                 205             


Glu Leu Arg Gln Arg Leu Ala Ala Arg Leu Glu Ala Leu Lys Glu Asn 
    210                 215                 220                 


Gly Gly Ala Arg Leu Ala Glu Tyr His Ala Lys Ala Thr Glu His Leu 
225                 230                 235                 240 


Ser Thr Leu Ser Glu Lys Ala Lys Pro Ala Leu Glu Asp Leu Arg Gln 
                245                 250                 255     


Gly Leu Leu Pro Val Leu Glu Ser Phe Lys Val Ser Phe Leu Ser Ala 
            260                 265                 270         


Leu Glu Glu Tyr Thr Lys Lys Leu Asn Thr Gln 
        275                 280             


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

<220>
<223>  protease cleavage site

<400>  5

Pro Ala Pro Ser Pro 
1               5   


<210>  6
<211>  4
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  protease cleavage site

<400>  6

Pro Pro Ser Pro 
1               


<210>  7
<211>  4
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  protease cleavage site

<400>  7

Pro Pro Ala Pro 
1               


<210>  8
<211>  4
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  protease cleavage site

<400>  8

Pro Pro Thr Pro 
1               


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

<220>
<223>  protease cleavage site

<400>  9

Pro Pro Gly Pro 
1               


<210>  10
<211>  6
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  enzymatic cleavage site

<400>  10

Pro Arg Pro Pro Thr Pro 
1               5       


<210>  11
<211>  7
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  enzymatic cleavage site

<400>  11

Val Val Ala Pro Pro Ala Pro 
1               5           


<210>  12
<211>  7
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  enzymatic cleavage site

<400>  12

Val Val Ala Pro Pro Ser Pro 
1               5           


<210>  13
<211>  7
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  enzymatic cleavage site

<400>  13

Val Val Ala Pro Pro Thr Pro 
1               5           


<210>  14
<211>  7
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  enzymatic cleavage site

<400>  14

Val Val Ala Pro Pro Gly Pro 
1               5           


<210>  15
<211>  6
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  enzymatic cleavage site

<400>  15

Ala Pro Pro Ala Ala Pro 
1               5       


<210>  16
<211>  6
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  enzymatic cleavage site

<400>  16

Pro Arg Pro Pro Ala Pro 
1               5       


<210>  17
<211>  6
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  enzymatic cleavage site

<400>  17

Pro Arg Pro Pro Ser Pro 
1               5       


<210>  18
<211>  6
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  enzymatic cleavage site

<400>  18

Pro Arg Pro Pro Gly Pro 
1               5       


<210>  19
<211>  9
<212>  PRT
<213>  Homo sapiens

<400>  19

Cys Asp Leu Pro Gln Thr His Ser Leu 
1               5                   


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

<220>
<223>  hexahistidine affinity tag

<400>  20

His His His His His His 
1               5       


<210>  21
<211>  195
<212>  PRT
<213>  Homo sapiens

<400>  21

Met Gly Lys Ile Ser Ser Leu Pro Thr Gln Leu Phe Lys Cys Cys Phe 
1               5                   10                  15      


Cys Asp Phe Leu Lys Val Lys Met His Thr Met Ser Ser Ser His Leu 
            20                  25                  30          


Phe Tyr Leu Ala Leu Cys Leu Leu Thr Phe Thr Ser Ser Ala Thr Ala 
        35                  40                  45              


Gly Pro Glu Thr Leu Cys Gly Ala Glu Leu Val Asp Ala Leu Gln Phe 
    50                  55                  60                  


Val Cys Gly Asp Arg Gly Phe Tyr Phe Asn Lys Pro Thr Gly Tyr Gly 
65                  70                  75                  80  


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


Phe Arg Ser Cys Asp Leu Arg Arg Leu Glu Met Tyr Cys Ala Pro Leu 
            100                 105                 110         


Lys Pro Ala Lys Ser Ala Arg Ser Val Arg Ala Gln Arg His Thr Asp 
        115                 120                 125             


Met Pro Lys Thr Gln Lys Tyr Gln Pro Pro Ser Thr Asn Lys Asn Thr 
    130                 135                 140                 


Lys Ser Gln Arg Arg Lys Gly Trp Pro Lys Thr His Pro Gly Gly Glu 
145                 150                 155                 160 


Gln Lys Glu Gly Thr Glu Ala Ser Leu Gln Ile Arg Gly Lys Lys Lys 
                165                 170                 175     


Glu Gln Arg Arg Glu Ile Gly Ser Arg Asn Ala Glu Cys Arg Gly Lys 
            180                 185                 190         


Lys Gly Lys 
        195 


