                         SEQUENCE LISTING

<110>  Roche Diagnostics GmbH
       F. Hoffmann-La Roche AG
       Roche Diagnostics Operations, Inc.
 
<120>  Thermostable Affinity Polypeptides

<130>  RD36580PC

<150>  EP21180048.7
<151>  2021-06-17

<160>  19    

<170>  PatentIn version 3.5

<210>  1
<211>  115
<212>  PRT
<213>  Geobacillus thermoglucosidasius

<400>  1

Tyr Asn Lys Gly Ala Ile Gly Ile Ile Pro Lys Glu Ile Arg Asp Lys 
1               5                   10                  15      


Leu Val Gly Lys Lys Phe Asn Ser Phe Asp Asp Phe Arg Lys Glu Phe 
            20                  25                  30          


Trp Lys Cys Val Ala Asn Ser Ser Tyr Ala Lys Glu Phe Arg Ser Arg 
        35                  40                  45              


Asn Ile Ala Arg Met Leu Glu Gly Asn Ala Pro Ile Ala Pro Lys Ala 
    50                  55                  60                  


Glu His Tyr Gly Lys Leu Lys Ser Tyr Ile Leu His His Lys Gln Pro 
65                  70                  75                  80  


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


Ser Pro Arg Met His Gln Ile Ile Leu Asp Pro Ala Tyr Tyr Phe Gly 
            100                 105                 110         


Lys Lys Gly 
        115 


<210>  2
<211>  69
<212>  PRT
<213>  Geobacillus thermoglucosidasius

<400>  2

Glu Asn Lys Leu Ser Lys Glu Glu Leu Ile Glu Leu Val Lys Lys Ile 
1               5                   10                  15      


Cys Asp Pro Lys Leu Ser Asp Glu Glu Val Ser Glu Tyr Ile Asp Ile 
            20                  25                  30          


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


Asn Asp Glu Glu Leu Ser Pro Glu Glu Ile Val Glu Ile Ala Leu Asn 
    50                  55                  60                  


Tyr Lys Asp Arg Asn 
65                  


<210>  3
<211>  115
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Gth colicin polypeptide mutein fragment

<400>  3

Tyr Asn Lys Gly Ala Ile Gly Ile Ile Pro Lys Glu Ile Arg Asp Lys 
1               5                   10                  15      


Leu Val Gly Lys Lys Phe Asn Ser Phe Asp Asp Phe Arg Lys Glu Phe 
            20                  25                  30          


Trp Lys Ala Val Ala Asn Ser Ser Tyr Ala Lys Glu Phe Arg Ser Arg 
        35                  40                  45              


Asn Ile Ala Arg Met Leu Glu Gly Asn Ala Pro Ile Ala Pro Lys Ala 
    50                  55                  60                  


Glu His Tyr Gly Lys Leu Lys Ser Tyr Ile Leu His Ala Lys Gln Pro 
65                  70                  75                  80  


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


Ser Pro Arg Met His Gln Ile Ile Leu Asp Pro Ala Tyr Tyr Phe Gly 
            100                 105                 110         


Lys Lys Gly 
        115 


<210>  4
<211>  69
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Gth immunity polypeptide mutein fragment

<400>  4

Glu Asn Lys Leu Ser Lys Glu Glu Leu Ile Glu Leu Val Lys Lys Ile 
1               5                   10                  15      


Ala Asp Pro Lys Leu Ser Asp Glu Glu Val Ser Glu Tyr Ile Asp Ile 
            20                  25                  30          


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


Asn Asp Glu Glu Leu Ser Pro Glu Glu Ile Val Glu Ile Ala Leu Asn 
    50                  55                  60                  


Tyr Lys Asp Arg Asn 
65                  


<210>  5
<211>  6
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  (His)6-tag

<400>  5

His His His His His His 
1               5       


<210>  6
<211>  8
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  (His)8-tag

<400>  6

His His His His His His His His 
1               5               


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

<220>
<223>  GGGS-linker

<400>  7

Gly Gly Gly Ser 
1               


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

<220>
<223>  (GGGS)5-Linker

<400>  8

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


Gly Gly Gly Ser 
            20  


<210>  9
<211>  12
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Linker

<400>  9

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


<210>  10
<211>  265
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Heavy Chain Fab <IGF-1>-GthCol mut

<400>  10

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


Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ala Phe Ser Asn Leu 
            20                  25                  30          


Trp Met Asn Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 
        35                  40                  45              


Gly Gln Ile Tyr Pro Gly Asp Gly Asp Thr Asn Tyr Tyr Gly Asp Phe 
    50                  55                  60                  


Lys Gly Lys Ala Thr Leu Thr Ala Asp Ser Ser Ser Ser Thr Ala Tyr 
65                  70                  75                  80  


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


Ala Arg Ile Gly Tyr Tyr Gly Met Asp Tyr Trp Gly Gln Gly Thr Ser 
            100                 105                 110         


Val Thr Val Ser Ser Gly Gly Ser Ser Gly Gly Ala Gly Ser Gly Gly 
        115                 120                 125             


Gly Gly Tyr Asn Lys Gly Ala Ile Gly Ile Ile Pro Lys Glu Ile Arg 
    130                 135                 140                 


Asp Lys Leu Val Gly Lys Lys Phe Asn Ser Phe Asp Asp Phe Arg Lys 
145                 150                 155                 160 


Glu Phe Trp Lys Ala Val Ala Asn Ser Ser Tyr Ala Lys Glu Phe Arg 
                165                 170                 175     


Ser Arg Asn Ile Ala Arg Met Leu Glu Gly Asn Ala Pro Ile Ala Pro 
            180                 185                 190         


Lys Ala Glu His Tyr Gly Lys Leu Lys Ser Tyr Ile Leu His Ala Lys 
        195                 200                 205             


Gln Pro Ile Glu Lys Gly Gly Glu Val Tyr Asn Leu Asp Asn Leu Ile 
    210                 215                 220                 


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


Phe Gly Lys Lys Gly Gly Gly Gly Tyr Arg Tyr Arg Gln Gly Gly Gly 
                245                 250                 255     


Ser His His His His His His His His 
            260                 265 


<210>  11
<211>  109
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Light Chain Fab <IGF-1>

<400>  11

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


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


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


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


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


<210>  12
<211>  219
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Heavy Chain Fab <IGF-1> -GthIM mut

<400>  12

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


Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ala Phe Ser Asn Leu 
            20                  25                  30          


Trp Met Asn Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 
        35                  40                  45              


Gly Gln Ile Tyr Pro Gly Asp Gly Asp Thr Asn Tyr Tyr Gly Asp Phe 
    50                  55                  60                  


Lys Gly Lys Ala Thr Leu Thr Ala Asp Ser Ser Ser Ser Thr Ala Tyr 
65                  70                  75                  80  


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


Ala Arg Ile Gly Tyr Tyr Gly Met Asp Tyr Trp Gly Gln Gly Thr Ser 
            100                 105                 110         


Val Thr Val Ser Ser Gly Gly Ser Ser Gly Gly Ala Gly Ser Gly Gly 
        115                 120                 125             


Gly Gly Glu Asn Lys Leu Ser Lys Glu Glu Leu Ile Glu Leu Val Lys 
    130                 135                 140                 


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


Asp Ile Leu Glu Lys Asn Val Pro His Pro Ala Pro Ser Asp Leu Ile 
                165                 170                 175     


Phe Trp Asn Asp Glu Glu Leu Ser Pro Glu Glu Ile Val Glu Ile Ala 
            180                 185                 190         


Leu Asn Tyr Lys Asp Arg Asn Gly Gly Gly Tyr Arg Tyr Arg Gln Gly 
        195                 200                 205             


Gly Gly Ser His His His His His His His His 
    210                 215                 


<210>  13
<211>  137
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Gth colicin mut + aux

<400>  13

Met Gly Tyr Asn Lys Gly Ala Ile Gly Ile Ile Pro Lys Glu Ile Arg 
1               5                   10                  15      


Asp Lys Leu Val Gly Lys Lys Phe Asn Ser Phe Asp Asp Phe Arg Lys 
            20                  25                  30          


Glu Phe Trp Lys Ala Val Ala Asn Ser Ser Tyr Ala Lys Glu Phe Arg 
        35                  40                  45              


Ser Arg Asn Ile Ala Arg Met Leu Glu Gly Asn Ala Pro Ile Ala Pro 
    50                  55                  60                  


Lys Ala Glu His Tyr Gly Lys Leu Lys Ser Tyr Ile Leu His Ala Lys 
65                  70                  75                  80  


Gln Pro Ile Glu Lys Gly Gly Glu Val Tyr Asn Leu Asp Asn Leu Ile 
                85                  90                  95      


Ile Thr Ser Pro Arg Met His Gln Ile Ile Leu Asp Pro Ala Tyr Tyr 
            100                 105                 110         


Phe Gly Lys Lys Gly Gly Gly Gly Tyr Arg Tyr Arg Gln Gly Gly Gly 
        115                 120                 125             


Ser His His His His His His His His 
    130                 135         


<210>  14
<211>  91
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Gth immunity mut. + aux

<400>  14

Met Gly Glu Asn Lys Leu Ser Lys Glu Glu Leu Ile Glu Leu Val Lys 
1               5                   10                  15      


Lys Ile Ala Asp Pro Lys Leu Ser Asp Glu Glu Val Ser Glu Tyr Ile 
            20                  25                  30          


Asp Ile Leu Glu Lys Asn Val Pro His Pro Ala Pro Ser Asp Leu Ile 
        35                  40                  45              


Phe Trp Asn Asp Glu Glu Leu Ser Pro Glu Glu Ile Val Glu Ile Ala 
    50                  55                  60                  


Leu Asn Tyr Lys Asp Arg Asn Gly Gly Gly Tyr Arg Tyr Arg Gln Gly 
65                  70                  75                  80  


Gly Gly Ser His His His His His His His His 
                85                  90      


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

<220>
<223>  transglutaminase peptide

<400>  15

Tyr Arg Tyr Arg Gln 
1               5   


<210>  16
<211>  115
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Gth colicin H77A

<400>  16

Tyr Asn Lys Gly Ala Ile Gly Ile Ile Pro Lys Glu Ile Arg Asp Lys 
1               5                   10                  15      


Leu Val Gly Lys Lys Phe Asn Ser Phe Asp Asp Phe Arg Lys Glu Phe 
            20                  25                  30          


Trp Lys Cys Val Ala Asn Ser Ser Tyr Ala Lys Glu Phe Arg Ser Arg 
        35                  40                  45              


Asn Ile Ala Arg Met Leu Glu Gly Asn Ala Pro Ile Ala Pro Lys Ala 
    50                  55                  60                  


Glu His Tyr Gly Lys Leu Lys Ser Tyr Ile Leu His Ala Lys Gln Pro 
65                  70                  75                  80  


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


Ser Pro Arg Met His Gln Ile Ile Leu Asp Pro Ala Tyr Tyr Phe Gly 
            100                 105                 110         


Lys Lys Gly 
        115 


<210>  17
<211>  4
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  repeating unit of poly(NANP)

<400>  17

Asn Ala Asn Pro 
1               


<210>  18
<211>  411
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Gth colicin mut + aux encoding

<400>  18
atgggctaca acaaaggtgc cattggcatt attccgaaag aaattcgtga taaactggtg       60

ggcaaaaaat tcaacagctt tgatgatttc cgcaaagagt tttggaaagc agttgcaaat      120

agcagctatg ccaaagaatt tcgtagccgt aatattgcac gtatgctgga aggtaatgca      180

ccgattgcac cgaaagcaga acattatggt aaactgaaaa gctatatcct gcatgccaaa      240

cagccgattg aaaaaggtgg tgaagtttat aatctggaca acctgattat taccagtccg      300

cgtatgcatc agattattct ggatccggca tattactttg gtaaaaaagg cggtggtggt      360

tatcgttatc gtcaaggtgg tggtagtcat catcaccacc atcaccatca c               411


<210>  19
<211>  273
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  GTH immunity mut + aux encoding

<400>  19
atgggcgaga acaaactgag caaagaagaa ctgattgagc tggtgaaaaa aatcgcagat       60

ccgaaactga gtgatgaaga agtgagcgag tatattgaca tcctggaaaa aaatgttccg      120

catccggcac cgagcgatct gattttttgg aacgatgaag aactgtcacc ggaagaaatt      180

gttgaaattg ccctgaacta taaagatcgc aatggtggtg gttatcgtta tcgtcaaggt      240

ggtggcagtc atcatcacca ccatcaccat cac                                   273


