                               SEQUENCE LISTING	

<110> AMGEN Research (MUNICH) GMBH
 
<120> B-Cell Depletion as a Diagnostic Marker

<130> AMG15257PCT


<150> 62/164,278
<151> 2015-05-20



<160> 22    

<170> PatentIn version 3.5

<210> 1
<211> 531
<212> PRT
<213> Artificial Sequence

<220>
<221>  MISC_FEATURE
<222>  (1)..(27)
<223>  leader sequence


<220>
<223> Synthetic CD19xCD3 bispecific single chain antibody 
      polypeptide

<400> 1
Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr Gly 
  1               5                  10                  15     

Val His Ser Asp Tyr Lys Asp Asp Asp Asp Lys Asp Ile Gln Leu Thr 
             20                  25                  30         

Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly Gln Arg Ala Thr Ile 
         35                  40                  45             

Ser Cys Lys Ala Ser Gln Ser Val Asp Tyr Asp Gly Asp Ser Tyr Leu 
     50                  55                  60                 

Asn Trp Tyr Gln Gln Ile Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr 
 65                  70                  75                  80 

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

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

Asp Ala Ala Thr Tyr His Cys Gln Gln Ser Thr Glu Asp Pro Trp Thr 
        115                 120                 125             

Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Gly Gly Gly Gly Ser Gly 
    130                 135                 140                 

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Gln Gln Ser 
145                 150                 155                 160 

Gly Ala Glu Leu Val Arg Pro Gly Ser Ser Val Lys Ile Ser Cys Lys 
                165                 170                 175     

Ala Ser Gly Tyr Ala Phe Ser Ser Tyr Trp Met Asn Trp Val Lys Gln 
            180                 185                 190         

Arg Pro Gly Gln Gly Leu Glu Trp Ile Gly Gln Ile Trp Pro Gly Asp 
        195                 200                 205             

Gly Asp Thr Asn Tyr Asn Gly Lys Phe Lys Gly Lys Ala Thr Leu Thr 
    210                 215                 220                 

Ala Asp Glu Ser Ser Ser Thr Ala Tyr Met Gln Leu Ser Ser Leu Ala 
225                 230                 235                 240 

Ser Glu Asp Ser Ala Val Tyr Phe Cys Ala Arg Arg Glu Thr Thr Thr 
                245                 250                 255     

Val Gly Arg Tyr Tyr Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr Thr 
            260                 265                 270         

Val Thr Val Ser Ser Gly Gly Gly Gly Ser Asp Ile Lys Leu Gln Gln 
        275                 280                 285             

Ser Gly Ala Glu Leu Ala Arg Pro Gly Ala Ser Val Lys Met Ser Cys 
    290                 295                 300                 

Lys Thr Ser Gly Tyr Thr Phe Thr Arg Tyr Thr Met His Trp Val Lys 
305                 310                 315                 320 

Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile Gly Tyr Ile Asn Pro Ser 
                325                 330                 335     

Arg Gly Tyr Thr Asn Tyr Asn Gln Lys Phe Lys Asp Lys Ala Thr Leu 
            340                 345                 350         

Thr Thr Asp Lys Ser Ser Ser Thr Ala Tyr Met Gln Leu Ser Ser Leu 
        355                 360                 365             

Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys Ala Arg Tyr Tyr Asp Asp 
    370                 375                 380                 

His Tyr Cys Leu Asp Tyr Trp Gly Gln Gly Thr Thr Leu Thr Val Ser 
385                 390                 395                 400 

Ser Val Glu Gly Gly Ser Gly Gly Ser Gly Gly Ser Gly Gly Ser Gly 
                405                 410                 415     

Gly Val Asp Asp Ile Gln Leu Thr Gln Ser Pro Ala Ile Met Ser Ala 
            420                 425                 430         

Ser Pro Gly Glu Lys Val Thr Met Thr Cys Arg Ala Ser Ser Ser Val 
        435                 440                 445             

Ser Tyr Met Asn Trp Tyr Gln Gln Lys Ser Gly Thr Ser Pro Lys Arg 
    450                 455                 460                 

Trp Ile Tyr Asp Thr Ser Lys Val Ala Ser Gly Val Pro Tyr Arg Phe 
465                 470                 475                 480 

Ser Gly Ser Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Ser Met 
                485                 490                 495     

Glu Ala Glu Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Ser Ser Asn 
            500                 505                 510         

Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys His His His 
        515                 520                 525             

His His His 
    530     

        


<210> 2
<211> 1596
<212> DNA
<213> Artificial Sequence

<220>
<223> Synthetic CD19xCD3 bispecific single chain antibody 
      polynucleotide

<400> 2
atgggatgga gctgtatcat cctcttcttg gtagcaacag ctacaggtgt ccactccgac       60

tacaaagatg atgacgataa ggatatccag ctgacccagt ctccagcttc tttggctgtg      120

tctctagggc agagggccac catctcctgc aaggccagcc aaagtgttga ttatgatggt      180

gatagttatt tgaactggta ccaacagatt ccaggacagc cacccaaact cctcatctat      240

gatgcatcca atctagtttc tgggatccca cccaggttta gtggcagtgg gtctgggaca      300

gacttcaccc tcaacatcca tcctgtggag aaggtggatg ctgcaaccta tcactgtcag      360

caaagtactg aggatccgtg gacgttcggt ggagggacca agctcgagat caaaggtggt      420

ggtggttctg gcggcggcgg ctccggtggt ggtggttctc aggtgcagct gcagcagtct      480

ggggctgagc tggtgaggcc tgggtcctca gtgaagattt cctgcaaggc ttctggctat      540

gcattcagta gctactggat gaactgggtg aagcagaggc ctggacaggg tcttgagtgg      600

attggacaga tttggcctgg agatggtgat actaactaca atggaaagtt caagggtaaa      660

gccactctga ctgcagacga atcctccagc acagcctaca tgcaactcag cagcctagca      720

tctgaggact ctgcggtcta tttctgtgca agacgggaga ctacgacggt aggccgttat      780

tactatgcta tggactactg gggccaaggg accacggtca ccgtctcctc cggaggtggt      840

ggatccgata tcaaactgca gcagtcaggg gctgaactgg caagacctgg ggcctcagtg      900

aagatgtcct gcaagacttc tggctacacc tttactaggt acacgatgca ctgggtaaaa      960

cagaggcctg gacagggtct ggaatggatt ggatacatta atcctagccg tggttatact     1020

aattacaatc agaagttcaa ggacaaggcc acattgacta cagacaaatc ctccagcaca     1080

gcctacatgc aactgagcag cctgacatct gaggactctg cagtctatta ctgtgcaaga     1140

tattatgatg atcattactg ccttgactac tggggccaag gcaccactct cacagtctcc     1200

tcagtcgaag gtggaagtgg aggttctggt ggaagtggag gttcaggtgg agtcgacgac     1260

attcagctga cccagtctcc agcaatcatg tctgcatctc caggggagaa ggtcaccatg     1320

acctgcagag ccagttcaag tgtaagttac atgaactggt accagcagaa gtcaggcacc     1380

tcccccaaaa gatggattta tgacacatcc aaagtggctt ctggagtccc ttatcgcttc     1440

agtggcagtg ggtctgggac ctcatactct ctcacaatca gcagcatgga ggctgaagat     1500

gctgccactt attactgcca acagtggagt agtaacccgc tcacgttcgg tgctgggacc     1560

aagctggagc tgaaacatca tcaccatcat cattag                               1596



<210> 3
<211> 124
<212> PRT
<213> Artificial Sequence

<220>
<223> Synthetic VH anti CD19 polypeptide

<400> 3
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 Ser Tyr 
            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 Trp Pro Gly Asp Gly Asp Thr Asn Tyr Asn Gly Lys Phe 
    50                  55                  60                  


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


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


Ala Arg Arg Glu Thr Thr Thr Val Gly Arg Tyr Tyr Tyr Ala Met Asp 
            100                 105                 110         


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


<210> 4
<211> 372
<212> DNA
<213> Artificial Sequence

<220>
<223> Synthetic VH anti CD19 polynucleotide

<400> 4
caggtgcagc tgcagcagtc tggggctgag ctggtgaggc ctgggtcctc agtgaagatt       60

tcctgcaagg cttctggcta tgcattcagt agctactgga tgaactgggt gaagcagagg      120

cctggacagg gtcttgagtg gattggacag atttggcctg gagatggtga tactaactac      180

aatggaaagt tcaagggtaa agccactctg actgcagacg aatcctccag cacagcctac      240

atgcaactca gcagcctagc atctgaggac tctgcggtct atttctgtgc aagacgggag      300

actacgacgg taggccgtta ttactatgct atggactact ggggccaagg gaccacggtc      360

accgtctcct cc                                                          372


<210> 5
<211> 111
<212> PRT
<213> Artificial Sequence

<220>
<223> Synthetic VL anti CD19 polypeptide

<400> 5
Asp Ile Gln Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly 
1               5                   10                  15      


Gln Arg Ala Thr Ile Ser Cys Lys Ala Ser Gln Ser Val Asp Tyr Asp 
            20                  25                  30          


Gly Asp Ser Tyr Leu Asn Trp Tyr Gln Gln Ile Pro Gly Gln Pro Pro 
        35                  40                  45              


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


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


Pro Val Glu Lys Val Asp Ala Ala Thr Tyr His Cys Gln Gln Ser Thr 
                85                  90                  95      


Glu Asp Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 
            100                 105                 110     


<210> 6
<211> 333
<212> DNA
<213> Artificial Sequence

<220>
<223> Synthetic VL anti CD19 polynucleotide

<400> 6
gatatccagc tgacccagtc tccagcttct ttggctgtgt ctctagggca gagggccacc       60

atctcctgca aggccagcca aagtgttgat tatgatggtg atagttattt gaactggtac      120

caacagattc caggacagcc acccaaactc ctcatctatg atgcatccaa tctagtttct      180

gggatcccac ccaggtttag tggcagtggg tctgggacag acttcaccct caacatccat      240

cctgtggaga aggtggatgc tgcaacctat cactgtcagc aaagtactga ggatccgtgg      300

acgttcggtg gagggaccaa gctcgagatc aaa                                   333


<210> 7
<211> 119
<212> PRT
<213> Artificial Sequence

<220>
<223> Synthetic VH anti CD3 polypeptide

<400> 7
Asp Ile Lys Leu Gln Gln Ser Gly Ala Glu Leu Ala Arg Pro Gly Ala 
1               5                   10                  15      


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


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


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


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


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


Ala Arg Tyr Tyr Asp Asp His Tyr Cys Leu Asp Tyr Trp Gly Gln Gly 
            100                 105                 110         


Thr Thr Leu Thr Val Ser Ser 
        115                 


<210> 8
<211> 357
<212> DNA
<213> Artificial Sequence

<220>
<223> Synthetic VH anti CD3 polynucleotide

<400> 8
gatatcaaac tgcagcagtc aggggctgaa ctggcaagac ctggggcctc agtgaagatg       60

tcctgcaaga cttctggcta cacctttact aggtacacga tgcactgggt aaaacagagg      120

cctggacagg gtctggaatg gattggatac attaatccta gccgtggtta tactaattac      180

aatcagaagt tcaaggacaa ggccacattg actacagaca aatcctccag cacagcctac      240

atgcaactga gcagcctgac atctgaggac tctgcagtct attactgtgc aagatattat      300

gatgatcatt actgccttga ctactggggc caaggcacca ctctcacagt ctcctca         357


<210> 9
<211> 106
<212> PRT
<213> Artificial Sequence

<220>
<223> Synthetic VL anti CD3 polypeptide

<400> 9
Asp Ile Gln Leu Thr Gln Ser Pro Ala 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 Tyr Met 
            20                  25                  30          


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


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


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


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


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


<210> 10
<211> 318
<212> DNA
<213> Artificial Sequence

<220>
<223> Synthetic polynucleotide

<400> 10
gacattcagc tgacccagtc tccagcaatc atgtctgcat ctccagggga gaaggtcacc       60

atgacctgca gagccagttc aagtgtaagt tacatgaact ggtaccagca gaagtcaggc      120

acctccccca aaagatggat ttatgacaca tccaaagtgg cttctggagt cccttatcgc      180

ttcagtggca gtgggtctgg gacctcatac tctctcacaa tcagcagcat ggaggctgaa      240

gatgctgcca cttattactg ccaacagtgg agtagtaacc cgctcacgtt cggtgctggg      300

accaagctgg agctgaaa                                                    318


<210> 11
<211> 15
<212> PRT
<213> Artificial Sequence

<220>
<223> Synthetic anti-CD19 L1 peptide

<400> 11
Lys Ala Ser Gln Ser Val Asp Tyr Asp Gly Asp Ser Tyr Leu Asn 
1               5                   10                  15  


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

<220>
<223> Synthetic anti-CD19 L2 peptide

<400> 12
Asp Ala Ser Asn Leu Val Ser 
1               5           


<210> 13
<211> 9
<212> PRT
<213> Artificial Sequence

<220>
<223> Synthetic anti-CD19 L3 peptide

<400> 13
Gln Gln Ser Thr Glu Asp Pro Trp Thr 
1               5                   


<210> 14
<211> 5
<212> PRT
<213> Artificial Sequence

<220>
<223> Synthetic anti-CD19 H1 peptide

<400> 14
Ser Tyr Trp Met Asn 
1               5   


<210> 15
<211> 17
<212> PRT
<213> Artificial Sequence

<220>
<223> Synthetic anti-CD19 H2 peptide

<400> 15
Gln Ile Trp Pro Gly Asp Gly Asp Thr Asn Tyr Asn Gly Lys Phe Lys 
1               5                   10                  15      


Gly 
    


<210> 16
<211> 15
<212> PRT
<213> Artificial Sequence

<220>
<223> Synthetic anti-CD19 H3 peptide

<400> 16
Arg Glu Thr Thr Thr Val Gly Arg Tyr Tyr Tyr Ala Met Asp Tyr 
1               5                   10                  15  


<210> 17
<211> 5
<212> PRT
<213> Artificial Sequence

<220>
<223> Synthetic anti-CD3 H1 peptide

<400> 17
Arg Tyr Thr Met His 
1               5   


<210> 18
<211> 17
<212> PRT
<213> Artificial Sequence

<220>
<223> Synthetic anti-CD3 H2 peptide

<400> 18
Tyr Ile Asn Pro Ser Arg Gly Tyr Thr Asn Tyr Asn Gln Lys Phe Lys 
1               5                   10                  15      


Asp 
    


<210> 19
<211> 10
<212> PRT
<213> Artificial Sequence

<220>
<223> Synthetic anti-CD3 H3 peptide

<400> 19
Tyr Tyr Asp Asp His Tyr Cys Leu Asp Tyr 
1               5                   10  


<210> 20
<211> 10
<212> PRT
<213> Artificial Sequence

<220>
<223> Synthetic anti-CD3 L1 peptide

<400> 20
Arg Ala Ser Ser Ser Val Ser Tyr Met Asn 
1               5                   10  


<210> 21
<211> 7
<212> PRT
<213> Artificial Sequence

<220>
<223> Synthetic anti-CD3 L2 peptide

<400> 21
Asp Thr Ser Lys Val Ala Ser 
1               5           


<210> 22
<211> 9
<212> PRT
<213> Artificial Sequence

<220>
<223> Synthetic anti-CD3 L3 peptide

<400> 22
Gln Gln Trp Ser Ser Asn Pro Leu Thr 
1               5                   


