                         SEQUENCE LISTING

<110>  Roche Diagnostics GmbH
       F. Hoffmann-La Roche AG
 
<120>  Detection of a posttranslationally modified polypeptide by a 
       bi-valent binding agent

<130>  27204 WO-WN

<150>  EP10196687.7
<151>  2010-12-23

<150>  EP11173832.4
<151>  2011-07-13

<160>  26    

<170>  PatentIn version 3.5

<210>  1
<211>  122
<212>  PRT
<213>  Mus musculus

<400>  1

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


Gly Gly Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser 
            20                  25                  30          


Asp Tyr Pro Met Ser Trp Val Arg Gln Thr Pro Glu Lys Arg Leu Glu 
        35                  40                  45              


Trp Val Ala Thr Ile Thr Thr Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp 
    50                  55                  60                  


Ser Ile Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr 
65                  70                  75                  80  


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


Tyr Cys Thr Arg Val Lys Thr Asp Leu Trp Trp Gly Leu Ala Tyr Trp 
            100                 105                 110         


Gly Gln Gly Thr Leu Val Thr Val Ser Ala 
        115                 120         


<210>  2
<211>  116
<212>  PRT
<213>  Mus musculus

<400>  2

Gln Leu Val Leu Thr Gln Ser Ser Ser Ala Ser Phe Ser Leu Gly Ala 
1               5                   10                  15      


Ser Ala Lys Leu Thr Cys Thr Leu Ser Ser Gln His Ser Thr Tyr Thr 
            20                  25                  30          


Ile Glu Trp Tyr Gln Gln Gln Pro Leu Lys Pro Pro Lys Tyr Val Met 
        35                  40                  45              


Glu Leu Lys Lys Asp Gly Ser His Thr Thr Gly Asp Gly Ile Pro Asp 
    50                  55                  60                  


Arg Phe Ser Gly Ser Ser Ser Gly Ala Asp Arg Tyr Leu Ser Ile Ser 
65                  70                  75                  80  


Asn Ile Gln Pro Glu Asp Glu Ser Ile Tyr Ile Cys Gly Val Gly Asp 
                85                  90                  95      


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


Thr Val Leu Gly 
        115     


<210>  3
<211>  121
<212>  PRT
<213>  Mus musculus

<400>  3

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


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


Val Ile His Trp Val Lys Gln Lys Pro Gly Gln Gly Leu Glu Trp Ile 
        35                  40                  45              


Gly Tyr Leu Asn Pro Tyr Asn Asp Asn Thr Lys Tyr Asn Glu Lys Phe 
    50                  55                  60                  


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


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


Ala Arg Arg Gly Ile Tyr Ala Tyr Asp His Tyr Phe Asp Tyr Trp Gly 
            100                 105                 110         


Gln Gly Thr Ser Leu Thr Val Ser Ser 
        115                 120     


<210>  4
<211>  106
<212>  PRT
<213>  Mus musculus

<400>  4

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


Glu Lys Val Thr Leu Thr Cys Ser Ala Ser Ser Ser Val Asn Tyr Met 
            20                  25                  30          


Tyr Trp Tyr Gln Gln Lys Pro Gly Ser Ser Pro Arg Leu Leu Ile Tyr 
        35                  40                  45              


Asp Thr Ser Asn Leu Ala Ser Gly Val Pro Val Arg Phe Ser Gly Ser 
    50                  55                  60                  


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


Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Ser Thr Tyr Pro Leu Thr 
                85                  90                  95      


Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys 
            100                 105     


<210>  5
<211>  19
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  19mer ssDNA

<400>  5
agtctattaa tgcttctgc                                                    19


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

<220>
<223>  17mer ssDNA

<400>  6
agttctatcg tcgtcca                                                      17


<210>  7
<211>  19
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  complementary 19mer ssDNA

<400>  7
gcagaagcat taatagact                                                    19


<210>  8
<211>  17
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  complementary 17mer ssDNA

<400>  8
tggacgacga tagaact                                                      17


<210>  9
<211>  33
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic peptide


<220>
<221>  MOD_RES
<222>  (16)..(17)
<223>  bAla

<400>  9

Glu Arg Ala Glu Gln Gln Arg Ile Arg Ala Glu Arg Glu Lys Glu Xaa 
1               5                   10                  15      


Xaa Ser Leu Lys Asp Arg Ile Glu Lys Arg Arg Arg Ala Glu Arg Ala 
            20                  25                  30          


Glu 
    


<210>  10
<211>  32
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic peptide


<220>
<221>  MOD_RES
<222>  (16)..(17)
<223>  Amino-trioxa-octanoic-acid (X represents 
       Amino-trioxa-octanoic-acid)

<400>  10

Ser Leu Lys Asp Arg Ile Glu Arg Arg Arg Ala Glu Arg Ala Glu Xaa 
1               5                   10                  15      


Xaa Glu Arg Ala Glu Gln Gln Arg Ile Arg Ala Glu Arg Glu Lys Glu 
            20                  25                  30          


<210>  11
<211>  27
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic peptide

<400>  11

Phe Asp Glu Arg Gln Pro Tyr Ala His Met Asn Gly Gly Arg Lys Asn 
1               5                   10                  15      


Glu Arg Ala Leu Pro Leu Pro Gln Ser Ser Thr 
            20                  25          


<210>  12
<211>  28
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic peptide

<400>  12

Tyr Glu Glu His Ile Pro Tyr Thr His Met Asn Gly Gly Lys Lys Asn 
1               5                   10                  15      


Gly Arg Ile Leu Thr Leu Pro Arg Ser Asn Pro Ser 
            20                  25              


<210>  13
<211>  5
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic peptide (Spacer)

<400>  13

Gly Gly Gly Gly Ser 
1               5   


<210>  14
<211>  9
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  HA-Tag

<400>  14

Tyr Pro Tyr Asp Val Pro Asp Tyr Ala 
1               5                   


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

<220>
<223>  Avi-Tag

<400>  15

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


<210>  16
<211>  10
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Sortase cleavage Tag

<400>  16

Leu Pro Glu Thr Gly Gly Gly Ser Gly Ser 
1               5                   10  


<210>  17
<211>  26
<212>  PRT
<213>  Homo sapiens


<220>
<221>  misc_feature
<223>  human HER3 (1242-1267)

<400>  17

Pro Leu His Pro Val Pro Ile Met Pro Thr Ala Gly Thr Thr Pro Asp 
1               5                   10                  15      


Glu Asp Tyr Glu Tyr Met Asn Arg Gln Arg 
            20                  25      


<210>  18
<211>  13
<212>  PRT
<213>  Homo sapiens


<220>
<221>  misc_feature
<223>  human HER3 (1283-1295)

<400>  18

Pro Ala Ser Glu Gln Gly Tyr Glu Glu Met Arg Ala Phe 
1               5                   10              


<210>  19
<211>  120
<212>  PRT
<213>  Mus musculus


<220>
<221>  misc_feature
<223>  VH (mAb 30.4.33)

<400>  19

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


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


Ile Ile His Trp Val Lys Gln Arg Pro Gly Gln Asp Leu Glu Trp Ile 
        35                  40                  45              


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


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


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


Thr Arg His Gly Tyr Tyr Arg Ser Asp Gly Phe Asp Tyr Trp Gly Gln 
            100                 105                 110         


Gly Thr Thr Leu Thr Val Ser Ser 
        115                 120 


<210>  20
<211>  108
<212>  PRT
<213>  Mus musculus


<220>
<221>  misc_feature
<223>  VL (mAb 30.4.33)

<400>  20

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


Glu Lys Val Thr Met Thr Cys Arg Ala Ser Ser Ser Val Ser Ser Ser 
            20                  25                  30          


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


Ile Tyr Ser Thr Ser Thr Leu Ala Ser Gly Val Pro Ala Arg Phe Ser 
    50                  55                  60                  


Gly Ser Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Gly Val Glu 
65                  70                  75                  80  


Thr Glu Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Tyr Gly Thr Ser Pro 
                85                  90                  95      


Tyr Thr Phe Gly Ser Gly Thr Lys Val Asp Ile Lys 
            100                 105             


<210>  21
<211>  120
<212>  PRT
<213>  Mus musculus


<220>
<221>  misc_feature
<223>  VH (mAb 7.2.32)

<400>  21

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


Lys Gly Ser Leu Gln Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asn 
            20                  25                  30          


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


Trp Val Ala Arg Ile Arg Thr Glu Ser Ser Asp Tyr Ala Thr Asp Tyr 
    50                  55                  60                  


Ala Asp Ser Val Lys Asp Arg Phe Ile Ile Ser Arg Asp Asp Ser Gln 
65                  70                  75                  80  


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


Ile Tyr Tyr Cys Val Arg Ser Ser Gly Phe Asp Tyr Trp Gly Gln Gly 
            100                 105                 110         


Thr Thr Leu Thr Val Ser Ser Ser 
        115                 120 


<210>  22
<211>  110
<212>  PRT
<213>  Mus musculus


<220>
<221>  misc_feature
<223>  VL (mAb 7.2.32)

<400>  22

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


Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Val His Asp Asn 
            20                  25                  30          


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


Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro Asp 
    50                  55                  60                  


Arg Phe Gly Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile Ser 
65                  70                  75                  80  


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


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


<210>  23
<211>  123
<212>  PRT
<213>  Mus musculus


<220>
<221>  misc_feature
<223>  VH (mAb 4.1.15)

<400>  23

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


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


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


Trp Ile Gly Arg Ile Phe Pro Gly Asn Gly Asp Thr Asp Tyr Asn Gly 
    50                  55                  60                  


Asn Phe Arg Ala Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr 
65                  70                  75                  80  


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


Phe Cys Ala Arg Ser Arg Gly Leu Arg Gln Gly Ala Gly Phe Ala Tyr 
            100                 105                 110         


Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ala 
        115                 120             


<210>  24
<211>  113
<212>  PRT
<213>  Mus musculus


<220>
<221>  misc_feature
<223>  VL (mAb 4.1.15)

<400>  24

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


Gln Lys Ala Thr Met Ser Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser 
            20                  25                  30          


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


Ser Pro Lys Leu Leu Val Tyr Phe Ala Ser Thr Arg Glu Ser Gly Val 
    50                  55                  60                  


Pro Asp Arg Phe Ile Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 
65                  70                  75                  80  


Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Ala Tyr Phe Cys Gln Gln 
                85                  90                  95      


His Tyr Ser Asn Pro Arg Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile 
            100                 105                 110         


Lys 
    


<210>  25
<211>  18
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  17mer-Oligo


<220>
<221>  misc_feature
<222>  (1)..(1)
<223>  N = Gly-Gly-Aminolinker-(Spacer C3)3

<220>
<221>  modified_base
<222>  (18)..(18)
<223>  'A' is fluorescein labeled

<400>  25
nagttctatc gtcgtcca                                                     18


<210>  26
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  19mer-Oligo


<220>
<221>  modified_base
<222>  (1)..(1)
<223>  'A' is fluorescein labeled

<220>
<221>  misc_feature
<222>  (20)..(20)
<223>  N = (Spacer C3)3-Aminolinker-Gly-Gly

<400>  26
agtctattaa tgcttctgcn                                                   20


