                         SEQUENCE LISTING

<110>  Roche Diagnostics GmbH
       F. Hoffmann-La Roche AG
 
<120>  Chaperone-chaperone fusion polypeptides for reduction of 
       interferences and stabilization of immunoassays

<130>  31185 WO-IR

<150>  EP12006298.9 - 2403
<151>  2012-09-06

<160>  9     

<170>  PatentIn version 3.5

<210>  1
<211>  529
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Skp tandem SlyD

<400>  1

Ala Asp Lys Ile Ala Ile Val Asn Met Gly Ser Leu Phe Gln Gln Val 
1               5                   10                  15      


Ala Gln Lys Thr Gly Val Ser Asn Thr Leu Glu Asn Glu Phe Arg Gly 
            20                  25                  30          


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


Lys Lys Leu Gln Ser Met Lys Ala Gly Ser Asp Arg Thr Lys Leu Glu 
    50                  55                  60                  


Lys Asp Val Met Ala Gln Arg Gln Thr Phe Ala Gln Lys Ala Gln Ala 
65                  70                  75                  80  


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


Val Thr Arg Ile Gln Thr Ala Val Lys Ser Val Ala Asn Ser Gln Asp 
            100                 105                 110         


Ile Asp Leu Val Val Asp Ala Asn Ala Val Ala Tyr Asn Ser Ser Asp 
        115                 120                 125             


Val Lys Asp Ile Thr Ala Asp Val Leu Lys Gln Val Lys Gly Gly Gly 
    130                 135                 140                 


Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly 
145                 150                 155                 160 


Ser Gly Gly Gly Met Lys Val Ala Lys Asp Leu Val Val Ser Leu Ala 
                165                 170                 175     


Tyr Gln Val Arg Thr Glu Asp Gly Val Leu Val Asp Glu Ser Pro Val 
            180                 185                 190         


Ser Ala Pro Leu Asp Tyr Leu His Gly His Gly Ser Leu Ile Ser Gly 
        195                 200                 205             


Leu Glu Thr Ala Leu Glu Gly His Glu Val Gly Asp Lys Phe Asp Val 
    210                 215                 220                 


Ala Val Gly Ala Asn Asp Ala Tyr Gly Gln Tyr Asp Glu Asn Leu Val 
225                 230                 235                 240 


Gln Arg Val Pro Lys Asp Val Phe Met Gly Val Asp Glu Leu Gln Val 
                245                 250                 255     


Gly Met Arg Phe Leu Ala Glu Thr Asp Gln Gly Pro Val Pro Val Glu 
            260                 265                 270         


Ile Thr Ala Val Glu Asp Asp His Val Val Val Asp Gly Asn His Met 
        275                 280                 285             


Leu Ala Gly Gln Asn Leu Lys Phe Asn Val Glu Val Val Ala Ile Arg 
    290                 295                 300                 


Glu Ala Thr Glu Glu Glu Leu Ala His Gly His Val His Gly Ala His 
305                 310                 315                 320 


Asp His His His Asp His Asp His Asp Gly Gly Gly Ser Gly Gly Gly 
                325                 330                 335     


Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly 
            340                 345                 350         


Met Lys Val Ala Lys Asp Leu Val Val Ser Leu Ala Tyr Gln Val Arg 
        355                 360                 365             


Thr Glu Asp Gly Val Leu Val Asp Glu Ser Pro Val Ser Ala Pro Leu 
    370                 375                 380                 


Asp Tyr Leu His Gly His Gly Ser Leu Ile Ser Gly Leu Glu Thr Ala 
385                 390                 395                 400 


Leu Glu Gly His Glu Val Gly Asp Lys Phe Asp Val Ala Val Gly Ala 
                405                 410                 415     


Asn Asp Ala Tyr Gly Gln Tyr Asp Glu Asn Leu Val Gln Arg Val Pro 
            420                 425                 430         


Lys Asp Val Phe Met Gly Val Asp Glu Leu Gln Val Gly Met Arg Phe 
        435                 440                 445             


Leu Ala Glu Thr Asp Gln Gly Pro Val Pro Val Glu Ile Thr Ala Val 
    450                 455                 460                 


Glu Asp Asp His Val Val Val Asp Gly Asn His Met Leu Ala Gly Gln 
465                 470                 475                 480 


Asn Leu Lys Phe Asn Val Glu Val Val Ala Ile Arg Glu Ala Thr Glu 
                485                 490                 495     


Glu Glu Leu Ala His Gly His Val His Gly Ala His Asp His His His 
            500                 505                 510         


Asp His Asp His Asp Gly Gly Gly Ser His His His His His His His 
        515                 520                 525             


His 
    


<210>  2
<211>  161
<212>  PRT
<213>  escherichia coli

<400>  2

Met Lys Lys Trp Leu Leu Ala Ala Gly Leu Gly Leu Ala Leu Ala Thr 
1               5                   10                  15      


Ser Ala Gln Ala Ala Asp Lys Ile Ala Ile Val Asn Met Gly Ser Leu 
            20                  25                  30          


Phe Gln Gln Val Ala Gln Lys Thr Gly Val Ser Asn Thr Leu Glu Asn 
        35                  40                  45              


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


Gln Ala Lys Met Lys Lys Leu Gln Ser Met Lys Ala Gly Ser Asp Arg 
65                  70                  75                  80  


Thr Lys Leu Glu Lys Asp Val Met Ala Gln Arg Gln Thr Phe Ala Gln 
                85                  90                  95      


Lys Ala Gln Ala Phe Glu Gln Asp Arg Ala Arg Arg Ser Asn Glu Glu 
            100                 105                 110         


Arg Gly Lys Leu Val Thr Arg Ile Gln Thr Ala Val Lys Ser Val Ala 
        115                 120                 125             


Asn Ser Gln Asp Ile Asp Leu Val Val Asp Ala Asn Ala Val Ala Tyr 
    130                 135                 140                 


Asn Ser Ser Asp Val Lys Asp Ile Thr Ala Asp Val Leu Lys Gln Val 
145                 150                 155                 160 


Lys 
    


<210>  3
<211>  196
<212>  PRT
<213>  escherichia coli

<400>  3

Met Lys Val Ala Lys Asp Leu Val Val Ser Leu Ala Tyr Gln Val Arg 
1               5                   10                  15      


Thr Glu Asp Gly Val Leu Val Asp Glu Ser Pro Val Ser Ala Pro Leu 
            20                  25                  30          


Asp Tyr Leu His Gly His Gly Ser Leu Ile Ser Gly Leu Glu Thr Ala 
        35                  40                  45              


Leu Glu Gly His Glu Val Gly Asp Lys Phe Asp Val Ala Val Gly Ala 
    50                  55                  60                  


Asn Asp Ala Tyr Gly Gln Tyr Asp Glu Asn Leu Val Gln Arg Val Pro 
65                  70                  75                  80  


Lys Asp Val Phe Met Gly Val Asp Glu Leu Gln Val Gly Met Arg Phe 
                85                  90                  95      


Leu Ala Glu Thr Asp Gln Gly Pro Val Pro Val Glu Ile Thr Ala Val 
            100                 105                 110         


Glu Asp Asp His Val Val Val Asp Gly Asn His Met Leu Ala Gly Gln 
        115                 120                 125             


Asn Leu Lys Phe Asn Val Glu Val Val Ala Ile Arg Glu Ala Thr Glu 
    130                 135                 140                 


Glu Glu Leu Ala His Gly His Val His Gly Ala His Asp His His His 
145                 150                 155                 160 


Asp His Asp His Asp Gly Cys Cys Gly Gly His Gly His Asp His Gly 
                165                 170                 175     


His Glu His Gly Gly Glu Gly Cys Cys Gly Gly Lys Gly Asn Gly Gly 
            180                 185                 190         


Cys Gly Cys His 
        195     


<210>  4
<211>  23
<212>  PRT
<213>  artificial sequence

<220>
<223>  Glycine-rich spacer

<400>  4

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


Gly Gly Gly Ser Gly Gly Gly 
            20              


<210>  5
<211>  12
<212>  PRT
<213>  artificial sequence

<220>
<223>  octa-histidine tag

<400>  5

Gly Gly Gly Ser His His His His His His His His 
1               5                   10          


<210>  6
<211>  270
<212>  PRT
<213>  escherichia coli

<400>  6

Met Lys Ser Leu Phe Lys Val Thr Leu Leu Ala Thr Thr Met Ala Val 
1               5                   10                  15      


Ala Leu His Ala Pro Ile Thr Phe Ala Ala Glu Ala Ala Lys Pro Ala 
            20                  25                  30          


Thr Ala Ala Asp Ser Lys Ala Ala Phe Lys Asn Asp Asp Gln Lys Ser 
        35                  40                  45              


Ala Tyr Ala Leu Gly Ala Ser Leu Gly Arg Tyr Met Glu Asn Ser Leu 
    50                  55                  60                  


Lys Glu Gln Glu Lys Leu Gly Ile Lys Leu Asp Lys Asp Gln Leu Ile 
65                  70                  75                  80  


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


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


Ala Gln Ala Lys Met Glu Lys Asp Ala Ala Asp Asn Glu Ala Lys Gly 
        115                 120                 125             


Lys Glu Tyr Arg Glu Lys Phe Ala Lys Glu Lys Gly Val Lys Thr Ser 
    130                 135                 140                 


Ser Thr Gly Leu Val Tyr Gln Val Val Glu Ala Gly Lys Gly Glu Ala 
145                 150                 155                 160 


Pro Lys Asp Ser Asp Thr Val Val Val Asn Tyr Lys Gly Thr Leu Ile 
                165                 170                 175     


Asp Gly Lys Glu Phe Asp Asn Ser Tyr Thr Arg Gly Glu Pro Leu Ser 
            180                 185                 190         


Phe Arg Leu Asp Gly Val Ile Pro Gly Trp Thr Glu Gly Leu Lys Asn 
        195                 200                 205             


Ile Lys Lys Gly Gly Lys Ile Lys Leu Val Ile Pro Pro Glu Leu Ala 
    210                 215                 220                 


Tyr Gly Lys Ala Gly Val Pro Gly Ile Pro Pro Asn Ser Thr Leu Val 
225                 230                 235                 240 


Phe Asp Val Glu Leu Leu Asp Val Lys Pro Ala Pro Lys Ala Asp Ala 
                245                 250                 255     


Lys Pro Glu Ala Asp Ala Lys Ala Ala Asp Ser Ala Lys Lys 
            260                 265                 270 


<210>  7
<211>  149
<212>  PRT
<213>  escherichia coli

<400>  7

Met Ser Glu Ser Val Gln Ser Asn Ser Ala Val Leu Val His Phe Thr 
1               5                   10                  15      


Leu Lys Leu Asp Asp Gly Thr Thr Ala Glu Ser Thr Arg Asn Asn Gly 
            20                  25                  30          


Lys Pro Ala Leu Phe Arg Leu Gly Asp Ala Ser Leu Ser Glu Gly Leu 
        35                  40                  45              


Glu Gln His Leu Leu Gly Leu Lys Val Gly Asp Lys Thr Thr Phe Ser 
    50                  55                  60                  


Leu Glu Pro Asp Ala Ala Phe Gly Val Pro Ser Pro Asp Leu Ile Gln 
65                  70                  75                  80  


Tyr Phe Ser Arg Arg Glu Phe Met Asp Ala Gly Glu Pro Glu Ile Gly 
                85                  90                  95      


Ala Ile Met Leu Phe Thr Ala Met Asp Gly Ser Glu Met Pro Gly Val 
            100                 105                 110         


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


Leu Ala Gly Gln Thr Val His Phe Asp Ile Glu Val Leu Glu Ile Asp 
    130                 135                 140                 


Pro Ala Leu Glu Ala 
145                 


<210>  8
<211>  199
<212>  PRT
<213>  pasteurella multocida

<400>  8

Met Lys Ile Ala Lys Asn Val Val Val Ser Ile Ala Tyr Gln Val Arg 
1               5                   10                  15      


Thr Glu Asp Gly Val Leu Val Asp Glu Ala Pro Val Asn Gln Pro Leu 
            20                  25                  30          


Glu Tyr Leu Gln Gly His Asn Asn Leu Val Ile Gly Leu Glu Asn Ala 
        35                  40                  45              


Leu Glu Gly Lys Ala Val Gly Asp Lys Phe Glu Val Arg Val Lys Pro 
    50                  55                  60                  


Glu Glu Ala Tyr Gly Glu Tyr Asn Glu Asn Met Val Gln Arg Val Pro 
65                  70                  75                  80  


Lys Asp Val Phe Gln Gly Val Asp Glu Leu Val Val Gly Met Arg Phe 
                85                  90                  95      


Ile Ala Asp Thr Asp Ile Gly Pro Leu Pro Val Val Ile Thr Glu Val 
            100                 105                 110         


Ala Glu Asn Asp Val Val Val Asp Gly Asn His Met Leu Ala Gly Gln 
        115                 120                 125             


Glu Leu Leu Phe Ser Val Glu Val Val Ala Thr Arg Glu Ala Thr Leu 
    130                 135                 140                 


Glu Glu Ile Ala His Gly His Ile His Gln Glu Gly Gly Cys Cys Gly 
145                 150                 155                 160 


Gly His His His Asp Ser Asp Glu Glu Gly His Gly Cys Gly Cys Gly 
                165                 170                 175     


Ser His His His His Glu His Glu His His Ala His Asp Gly Cys Cys 
            180                 185                 190         


Gly Asn Gly Gly Cys Lys His 
        195                 


<210>  9
<211>  487
<212>  PRT
<213>  Artificial

<220>
<223>  Skp SlpA SlpA

<400>  9

Ala Asp Lys Ile Ala Ile Val Asn Met Gly Ser Leu Phe Gln Gln Val 
1               5                   10                  15      


Ala Gln Lys Thr Gly Val Ser Asn Thr Leu Glu Asn Glu Phe Arg Gly 
            20                  25                  30          


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


Lys Lys Leu Gln Ser Met Lys Ala Gly Ser Asp Arg Thr Lys Leu Glu 
    50                  55                  60                  


Lys Asp Val Met Ala Gln Arg Gln Thr Phe Ala Gln Lys Ala Gln Ala 
65                  70                  75                  80  


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


Val Thr Arg Ile Gln Thr Ala Val Lys Ser Val Ala Asn Ser Gln Asp 
            100                 105                 110         


Ile Asp Leu Val Val Asp Ala Asn Ala Val Ala Tyr Asn Ser Ser Asp 
        115                 120                 125             


Val Lys Asp Ile Thr Ala Asp Val Leu Lys Gln Val Lys Gly Gly Gly 
    130                 135                 140                 


Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly 
145                 150                 155                 160 


Ser Gly Gly Gly Ser Glu Ser Val Gln Ser Asn Ser Ala Val Leu Val 
                165                 170                 175     


His Phe Thr Leu Lys Leu Asp Asp Gly Thr Thr Ala Glu Ser Thr Arg 
            180                 185                 190         


Asn Asn Gly Lys Pro Ala Leu Phe Arg Leu Gly Asp Ala Ser Leu Ser 
        195                 200                 205             


Glu Gly Leu Glu Gln His Leu Leu Gly Leu Lys Val Gly Asp Lys Thr 
    210                 215                 220                 


Thr Phe Ser Leu Glu Pro Asp Ala Ala Phe Gly Val Pro Ser Pro Asp 
225                 230                 235                 240 


Leu Ile Gln Tyr Phe Ser Arg Arg Glu Phe Met Asp Ala Gly Glu Pro 
                245                 250                 255     


Glu Ile Gly Ala Ile Met Leu Phe Thr Ala Met Asp Gly Ser Glu Met 
            260                 265                 270         


Pro Gly Val Ile Arg Glu Ile Asn Gly Asp Ser Ile Thr Val Asp Phe 
        275                 280                 285             


Asn His Pro Leu Ala Gly Gln Thr Val His Phe Asp Ile Glu Val Leu 
    290                 295                 300                 


Glu Ile Asp Pro Ala Leu Glu Gly Gly Gly Ser Gly Gly Gly Ser Gly 
305                 310                 315                 320 


Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Glu 
                325                 330                 335     


Ser Val Gln Ser Asn Ser Ala Val Leu Val His Phe Thr Leu Lys Leu 
            340                 345                 350         


Asp Asp Gly Thr Thr Ala Glu Ser Thr Arg Asn Asn Gly Lys Pro Ala 
        355                 360                 365             


Leu Phe Arg Leu Gly Asp Ala Ser Leu Ser Glu Gly Leu Glu Gln His 
    370                 375                 380                 


Leu Leu Gly Leu Lys Val Gly Asp Lys Thr Thr Phe Ser Leu Glu Pro 
385                 390                 395                 400 


Asp Ala Ala Phe Gly Val Pro Ser Pro Asp Leu Ile Gln Tyr Phe Ser 
                405                 410                 415     


Arg Arg Glu Phe Met Asp Ala Gly Glu Pro Glu Ile Gly Ala Ile Met 
            420                 425                 430         


Leu Phe Thr Ala Met Asp Gly Ser Glu Met Pro Gly Val Ile Arg Glu 
        435                 440                 445             


Ile Asn Gly Asp Ser Ile Thr Val Asp Phe Asn His Pro Leu Ala Gly 
    450                 455                 460                 


Gln Thr Val His Phe Asp Ile Glu Val Leu Glu Ile Asp Pro Ala Leu 
465                 470                 475                 480 


Glu His His His His His His 
                485         


