                         SEQUENCE LISTING

<110>  F. HOFFMANN-LA ROCHE
       INSTITUT PASTEUR
       CENTRE NATIONAL DE LA RECHERCHE SCIENTIFIQUE
       COMMISSARIAT A L'ENERGIE ATOMIQUE ET AUX ENERGIES 
       ALTERNATIVES
 
<120>  CAMELID SINGLE-DOMAIN ANTIBODY DIRECTED AGAINST AMYLOID BETA AND 
       METHODS FOR PRODUCING CONJUGATES THEREOF

<130>  XRN/SM/cc/F226-199

<150>  EP13306553.2
<151>  2013-11-13

<160>  18    

<170>  PatentIn version 3.5

<210>  1
<211>  13
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  CDR1 of VHH R3VE/Q

<400>  1

Ala Asp Ser Gly Ser Thr Phe Arg Asn Tyr Asn Ile Gly 
1               5                   10              


<210>  2
<211>  17
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  CDR2 of VHH R3VE/Q

<400>  2

Ala Val Ser Arg Thr Gly Ile Ser Thr His Val Ala Asp Ser Leu Gln 
1               5                   10                  15      


Gly 
    


<210>  3
<211>  18
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  CDR3 of VHH R3VE/Q

<400>  3

Ala Ala Gly Arg Pro Gly Val Gly Ala Val Asn Arg Ala Met Asp Tyr 
1               5                   10                  15      


Asp Tyr 
        


<210>  4
<211>  127
<212>  PRT
<213>  Lama pacos

<400>  4

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


Gly Asp Ser Leu Arg Leu Ser Cys Ala Asp Ser Gly Ser Thr Phe Arg 
            20                  25                  30          


Asn Tyr Asn Ile Gly Trp Phe Arg Gln Thr Pro Gly Gln Ala Arg Glu 
        35                  40                  45              


Phe Val Ala Ala Val Ser Arg Thr Gly Ile Ser Thr His Val Ala Asp 
    50                  55                  60                  


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


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


Ser Cys Ala Ala Gly Arg Pro Gly Val Gly Ala Val Asn Arg Ala Met 
            100                 105                 110         


Asp Tyr Asp Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 
        115                 120                 125         


<210>  5
<211>  127
<212>  PRT
<213>  Lama pacos

<400>  5

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


Gly Asp Ser Leu Arg Leu Ser Cys Ala Asp Ser Gly Ser Thr Phe Arg 
            20                  25                  30          


Asn Tyr Asn Ile Gly Trp Phe Arg Gln Thr Pro Gly Gln Ala Arg Glu 
        35                  40                  45              


Phe Val Ala Ala Val Ser Arg Thr Gly Ile Ser Thr His Val Ala Asp 
    50                  55                  60                  


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


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


Ser Cys Ala Ala Gly Arg Pro Gly Val Gly Ala Val Asn Arg Ala Met 
            100                 105                 110         


Asp Tyr Asp Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 
        115                 120                 125         


<210>  6
<211>  122
<212>  PRT
<213>  Lama pacos

<400>  6

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


Leu Arg Leu Ser Cys Ala Asp Ser Gly Ser Thr Phe Arg Asn Tyr Asn 
            20                  25                  30          


Ile Gly Trp Phe Arg Gln Thr Pro Gly Gln Ala Arg Glu Phe Val Ala 
        35                  40                  45              


Ala Val Ser Arg Thr Gly Ile Ser Thr His Val Ala Asp Ser Leu Gln 
    50                  55                  60                  


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


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


Ala Gly Arg Pro Gly Val Gly Ala Val Asn Arg Ala Met Asp Tyr Asp 
            100                 105                 110         


Tyr Trp Gly Gln Gly Thr Gln Val Thr Val 
        115                 120         


<210>  7
<211>  122
<212>  PRT
<213>  Lama pacos

<400>  7

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


Leu Arg Leu Ser Cys Ala Asp Ser Gly Ser Thr Phe Arg Asn Tyr Asn 
            20                  25                  30          


Ile Gly Trp Phe Arg Gln Thr Pro Gly Gln Ala Arg Glu Phe Val Ala 
        35                  40                  45              


Ala Val Ser Arg Thr Gly Ile Ser Thr His Val Ala Asp Ser Leu Gln 
    50                  55                  60                  


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


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


Ala Gly Arg Pro Gly Val Gly Ala Val Asn Arg Ala Met Asp Tyr Asp 
            100                 105                 110         


Tyr Trp Gly Gln Gly Thr Gln Val Thr Val 
        115                 120         


<210>  8
<211>  151
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  R3VQ-SH


<220>
<221>  DOMAIN
<222>  (5)..(6)
<223>  6xHis tag

<220>
<221>  DOMAIN
<222>  (14)..(19)
<223>  Thrombin cleavage site

<220>
<221>  DOMAIN
<222>  (23)..(144)
<223>  VHH R3VQ

<220>
<221>  DOMAIN
<222>  (145)..(148)
<223>  Spacer

<220>
<221>  DOMAIN
<222>  (150)..(151)
<223>  Spacer

<400>  8

Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 
1               5                   10                  15      


Arg Gly Ser Ala Ala Ala Val Gln Leu Gln Ala Ser Gly Gly Gly Leu 
            20                  25                  30          


Val Gln Thr Gly Asp Ser Leu Arg Leu Ser Cys Ala Asp Ser Gly Ser 
        35                  40                  45              


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


Ala Arg Glu Phe Val Ala Ala Val Ser Arg Thr Gly Ile Ser Thr His 
65                  70                  75                  80  


Val Ala Asp Ser Leu Gln Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala 
                85                  90                  95      


Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr 
            100                 105                 110         


Ala Val Tyr Ser Cys Ala Ala Gly Arg Pro Gly Val Gly Ala Val Asn 
        115                 120                 125             


Arg Ala Met Asp Tyr Asp Tyr Trp Gly Gln Gly Thr Gln Val Thr Val 
    130                 135                 140                 


Gly Gly Gly Ser Cys Ser Ala 
145                 150     


<210>  9
<211>  6
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  6xHis tag

<400>  9

His His His His His His 
1               5       


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

<220>
<223>  Thrombin cleavage site

<400>  10

Leu Val Pro Arg Gly Ser 
1               5       


<210>  11
<211>  4
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Spacer

<400>  11

Gly Gly Gly Ser 
1               


<210>  12
<211>  42
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Amyloid beta 42

<400>  12

Asp Ala Glu Phe Arg His Asp Ser Gly Tyr Glu Val His His Gln Lys 
1               5                   10                  15      


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


Gly Leu Met Val Gly Gly Val Val Ile Ala 
        35                  40          


<210>  13
<211>  41
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer

<400>  13
ggactagttg cggccgctgg ttgtggtttt ggtgtcttgg g                           41


<210>  14
<211>  123
<212>  PRT
<213>  Lama pacos

<400>  14

Met Ala Glu Val Gln Leu Gln Ala Ser Gly Gly Gly Leu Val Gln Ala 
1               5                   10                  15      


Gly Gly Ser Arg Arg Leu Ser Cys Ala Val Ser Gly Arg Thr Phe Ser 
            20                  25                  30          


Ser Tyr Asn Met Gly Trp Phe Arg Gln Ala Pro Gly Glu Glu Arg Glu 
        35                  40                  45              


Phe Val Ala Ala Ile Asn Trp Ser Ala Ser Thr Tyr Tyr Ala Asp Ser 
    50                  55                  60                  


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


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


Cys Asn Leu His Val Ala Thr Thr Ser Tyr Phe Gln Thr Ser Asp Tyr 
            100                 105                 110         


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


<210>  15
<211>  123
<212>  PRT
<213>  Lama pacos

<400>  15

Met Ala Glu Val Gln Leu Gln Ala Ser Gly Gly Gly Leu Val Gln Ala 
1               5                   10                  15      


Gly Gly Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Arg Thr Phe Ser 
            20                  25                  30          


Ser Tyr Asn Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu 
        35                  40                  45              


Phe Val Ala Ala Ile Asn Trp Ser Ala Ser Thr Tyr Tyr Ala Asp Ser 
    50                  55                  60                  


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


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


Cys Asn Leu His Val Ala Thr Thr Ser Tyr Phe Gln Thr Ser Asp Tyr 
            100                 105                 110         


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


<210>  16
<211>  124
<212>  PRT
<213>  Lama pacos

<400>  16

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


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


Ile Asn Val Met Gly Trp Tyr Arg Gln Ser Pro Asp Gly Val Arg Asp 
        35                  40                  45              


Leu Val Ala Thr Ile Thr Ala Asn Gly Val Thr Asn Tyr Ala Ala Ser 
    50                  55                  60                  


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


Phe Leu Gln Met Asn Ser Val Lys Pro Glu Asp Thr Ala Val Tyr Ile 
                85                  90                  95      


Cys Asn Val Glu Gly Glu Tyr Ser Gly Ser Tyr Val Ala Asp Phe Asp 
            100                 105                 110         


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


<210>  17
<211>  465
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence encoding R3VQ-SH

<400>  17
atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagcgcg       60

gccgcagtcc agctgcaggc gtctggagga ggattggtgc agactgggga ctctctgaga      120

ctctcctgtg cagactctgg aagtacattc agaaactata acataggctg gttccgccag      180

actccaggac aggcgcgtga atttgtcgca gctgttagtc ggacgggaat tagcacacac      240

gttgcagact ccctccaggg ccgattcacc atctccagag acaacgccaa gaacacggtg      300

tatctgcaaa tgaacagcct gaaacctgag gacacggccg tttattcctg tgcagcgggg      360

cgaccgggcg taggagctgt caacagggcg atggattatg actactgggg ccaggggacc      420

caggtcaccg tcggtggcgg atcctgttcc gcgtaataac tcgag                      465


<210>  18
<211>  152
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  R3VE-SH


<220>
<221>  misc_feature
<222>  (98)..(98)
<223>  Xaa can be any naturally occurring amino acid

<400>  18

Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 
1               5                   10                  15      


Arg Gly Ser Ala Ala Ala Glu Val Gln Leu Glu Ala Ser Gly Gly Gly 
            20                  25                  30          


Leu Val Gln Thr Gly Asp Ser Leu Arg Leu Ser Cys Ala Asp Ser Gly 
        35                  40                  45              


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


Gln Ala Arg Glu Phe Val Ala Ala Val Ser Arg Thr Gly Ile Ser Thr 
65                  70                  75                  80  


His Val Ala Asp Ser Leu Gln Gly Arg Phe Thr Ile Ser Arg Asp Asn 
                85                  90                  95      


Ala Xaa Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp 
            100                 105                 110         


Thr Ala Val Tyr Ser Cys Ala Ala Gly Arg Pro Gly Val Gly Ala Val 
        115                 120                 125             


Asn Arg Ala Met Asp Tyr Asp Tyr Trp Gly Gln Gly Thr Gln Val Thr 
    130                 135                 140                 


Val Gly Gly Gly Ser Cys Ser Ala 
145                 150         


