                         SEQUENCE LISTING
<110>  Chemotherapeutisches Forschungsinstitut Georg-Speyer-Haus
       and DRK-Blutspendedienst Baden-Wrttemberg - Hessen gGmbH
 
<120>  CAR-expressing NK-92 cells as cell therapeutic agents

<130>  C31927WO

<150>  EP 14 173 020.0
<151>  2014-06-18

<160>  16    

<170>  PatentIn version 3.5

<210>  1
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer

<400>  1
cttcagcagg gaccagaaac aa                                                22


<210>  2
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer

<400>  2
ccgcttaata ctgacgctct cg                                                22


<210>  3
<211>  19
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer

<400>  3
atcgccacgg cagaactca                                                    19


<210>  4
<211>  21
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer

<400>  4
gacccttcac ccaacgctta g                                                 21


<210>  5
<211>  24
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer

<400>  5
tcagtggaat gggcagcttc aagt                                              24


<210>  6
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer

<400>  6
ttcagcaagc cgagtcctgc gt                                                22


<210>  7
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer

<400>  7
actgataatt ccgtggtgtt gt                                                22


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

<220>
<223>  Primer

<400>  8
cactgtggct cactgctaga                                                   20


<210>  9
<211>  587
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Amplification product of CAR vector integration in TRAF2 gene

<400>  9
cttcagcagg gaccagaaac aaaactcaca ctctttcttc tctgagttga gactggaaaa       60

atgaaagatt gttttagggg aaacttgagg gaacagtctg ggcagcctgc agggcatggc      120

cctgttcctc cagggctggg aaagtcagca ctgctttctg gtggcgaact ggaagggcta      180

attcactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac      240

cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa      300

agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag      360

agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg      420

acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag      480

cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg      540

gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcgg                    587


<210>  10
<211>  503
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Amplification product of CAR vector integration in TRAF2 gene

<400>  10
atcgccacgg cagaactcat cgccgcctgc cttgcccgct gctggacagg ggctaggttg       60

ctgggcactg ataattccgt ggtgttgtcg aattcgatac tcgaggtcga ggcaattcga      120

gctcggtacc tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa      180

aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat ctgctttttg      240

cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag      300

ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc      360

gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa      420

tctctagcac cttccctctg cagctgctgg ctcagccgat tgtatatgct gggagctctg      480

cactaagcgt tgggtgaagg gtc                                              503


<210>  11
<211>  679
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Amplification product of CAR vector integration in intergenic 
       region of chromosome 2

<400>  11
tcagtggaat gggcagcttc aagttgatgt catttcaata gtaacttatt tcagtctaca       60

tacttcccaa gaatgcacca tctctttttt atgtatttat tattttgaga aagagtctca      120

ctctgtcgcc caggctggag tgcaatggca tgatcttggc tcactgtaac ctccgtctcc      180

tgggttcaag ccattctcct gtctcagcct cccgggtagt ggggttatag gcacacacca      240

ccacgcccgg ctaatttttg tatttttagt aaagatgggg tttcaccatg ttggccaggc      300

tgggctcaaa ctcttgactt caggtgatcc gcccaccttg gcctcccaaa gtgctgggat      360

gacaggcact ggaagggcta attcactccc aacgaagaca agatctgctt tttgcttgta      420

ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc      480

cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt      540

tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta      600

gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg      660

caggactcgg cttgctgaa                                                   679


<210>  12
<211>  376
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Amplification product of CAR vector integration in intergenic 
       region of chromosome 2

<400>  12
actgataatt ccgtggtgtt gtcgaattcg atactcgagg tcgaggcaat tcgagctcgg       60

tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa      120

aggggggact ggaagggcta attcactccc aacgaagaca agatctgctt tttgcttgta      180

ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc      240

cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt      300

tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta      360

gcagtgagcc acagtg                                                      376


<210>  13
<211>  241
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  ErbB2-specific scFv antibody fragment FRP5

<400>  13

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


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


Gly Met Asn Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Lys Trp Met 
        35                  40                  45              


Gly Trp Ile Asn Thr Ser Thr Gly Glu Ser Thr Phe Ala Asp Asp Phe 
    50                  55                  60                  


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


Leu Gln Ile Asn Asn Leu Lys Ser Glu Asp Ser Ala Thr Tyr Phe Cys 
                85                  90                  95      


Ala Arg Trp Glu Val Tyr His Gly Tyr Val Pro Tyr Trp Gly Gln Gly 
            100                 105                 110         


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


Ser Gly Gly Gly Gly Ser Asp Ile Gln Leu Thr Gln Ser His Lys Phe 
    130                 135                 140                 


Leu Ser Thr Ser Val Gly Asp Arg Val Ser Ile Thr Cys Lys Ala Ser 
145                 150                 155                 160 


Gln Asp Val Tyr Asn Ala Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln 
                165                 170                 175     


Ser Pro Lys Leu Leu Ile Tyr Ser Ala Ser Ser Arg Tyr Thr Gly Val 
            180                 185                 190         


Pro Ser Arg Phe Thr Gly Ser Gly Ser Gly Pro Asp Phe Thr Phe Thr 
        195                 200                 205             


Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Val Tyr Phe Cys Gln Gln 
    210                 215                 220                 


His Phe Arg Thr Pro Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile 
225                 230                 235                 240 


Lys 
    


<210>  14
<211>  723
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence encoding ErbB2-specific scFv antibody 
       fragment FRP5

<400>  14
caggtgcagc tgcagcagag cggccctgag ctgaagaagc ccggcgagac agtcaagatc       60

agctgcaagg ccagcggcta ccccttcacc aactacggca tgaactgggt gaaacaggcc      120

ccaggccagg gactgaagtg gatgggctgg atcaacacca gcaccggcga gagcaccttc      180

gccgacgact tcaagggcag attcgacttc agcctggaaa ccagcgccaa caccgcctac      240

ctgcagatca acaacctgaa gagcgaggac agcgccacct acttttgcgc cagatgggag      300

gtgtaccacg gctacgtgcc ctactggggc cagggcacca ccgtgaccgt gtccagcggc      360

ggagggggct ctggcggcgg aggatctggg ggagggggca gcgacatcca gctgacccag      420

agccacaagt ttctgagcac cagcgtgggc gaccgggtgt ccatcacctg caaagccagc      480

caggacgtgt acaacgccgt ggcctggtat cagcagaagc ctggccagag ccccaagctg      540

ctgatctaca gcgccagcag ccggtacacc ggcgtgccca gcaggttcac cggcagcggc      600

agcggcccag acttcacctt caccatcagc agcgtgcagg ccgaggacct ggccgtgtac      660

ttctgccagc agcacttccg gacccccttc accttcggct ccggcaccaa gctggaaatc      720

aag                                                                    723


<210>  15
<211>  504
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  ErbB2-specific chimeric antigen receptor

<400>  15

Met Asp Trp Ile Trp Arg Ile Leu Phe Leu Val Gly Ala Ala Thr Gly 
1               5                   10                  15      


Ala His Ser Gln Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Lys Lys 
            20                  25                  30          


Pro Gly Glu Thr Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Pro Phe 
        35                  40                  45              


Thr Asn Tyr Gly Met Asn Trp Val Lys Gln Ala Pro Gly Gln Gly Leu 
    50                  55                  60                  


Lys Trp Met Gly Trp Ile Asn Thr Ser Thr Gly Glu Ser Thr Phe Ala 
65                  70                  75                  80  


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


Thr Ala Tyr Leu Gln Ile Asn Asn Leu Lys Ser Glu Asp Ser Ala Thr 
            100                 105                 110         


Tyr Phe Cys Ala Arg Trp Glu Val Tyr His Gly Tyr Val Pro Tyr Trp 
        115                 120                 125             


Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly 
    130                 135                 140                 


Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Leu Thr Gln Ser 
145                 150                 155                 160 


His Lys Phe Leu Ser Thr Ser Val Gly Asp Arg Val Ser Ile Thr Cys 
                165                 170                 175     


Lys Ala Ser Gln Asp Val Tyr Asn Ala Val Ala Trp Tyr Gln Gln Lys 
            180                 185                 190         


Pro Gly Gln Ser Pro Lys Leu Leu Ile Tyr Ser Ala Ser Ser Arg Tyr 
        195                 200                 205             


Thr Gly Val Pro Ser Arg Phe Thr Gly Ser Gly Ser Gly Pro Asp Phe 
    210                 215                 220                 


Thr Phe Thr Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Val Tyr Phe 
225                 230                 235                 240 


Cys Gln Gln His Phe Arg Thr Pro Phe Thr Phe Gly Ser Gly Thr Lys 
                245                 250                 255     


Leu Glu Ile Lys Ala Leu Ser Asn Ser Ile Met Tyr Phe Ser His Phe 
            260                 265                 270         


Val Pro Val Phe Leu Pro Ala Lys Pro Thr Thr Thr Pro Ala Pro Arg 
        275                 280                 285             


Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg 
    290                 295                 300                 


Pro Glu Ala Ser Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly 
305                 310                 315                 320 


Leu Asp Lys Pro Phe Trp Val Leu Val Val Val Gly Gly Val Leu Ala 
                325                 330                 335     


Cys Tyr Ser Leu Leu Val Thr Val Ala Phe Ile Ile Phe Trp Val Arg 
            340                 345                 350         


Ser Lys Arg Ser Arg Leu Leu His Ser Asp Tyr Met Asn Met Thr Pro 
        355                 360                 365             


Arg Arg Pro Gly Pro Thr Arg Lys His Tyr Gln Pro Tyr Ala Pro Pro 
    370                 375                 380                 


Arg Asp Phe Ala Ala Tyr Arg Ser Arg Val Lys Phe Ser Arg Ser Ala 
385                 390                 395                 400 


Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu 
                405                 410                 415     


Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly 
            420                 425                 430         


Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu 
        435                 440                 445             


Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser 
    450                 455                 460                 


Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly 
465                 470                 475                 480 


Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu 
                485                 490                 495     


His Met Gln Ala Leu Pro Pro Arg 
            500                 


<210>  16
<211>  1515
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence encoding ErbB2-specific chimeric antigen 
       receptor

<400>  16
atggactgga tctggcggat tctgttcctg gtcggggctg ccacaggcgc ccacagccag       60

gtgcagctgc agcagagcgg ccctgagctg aagaagcccg gcgagacagt caagatcagc      120

tgcaaggcca gcggctaccc cttcaccaac tacggcatga actgggtgaa acaggcccca      180

ggccagggac tgaagtggat gggctggatc aacaccagca ccggcgagag caccttcgcc      240

gacgacttca agggcagatt cgacttcagc ctggaaacca gcgccaacac cgcctacctg      300

cagatcaaca acctgaagag cgaggacagc gccacctact tttgcgccag atgggaggtg      360

taccacggct acgtgcccta ctggggccag ggcaccaccg tgaccgtgtc cagcggcgga      420

gggggctctg gcggcggagg atctggggga gggggcagcg acatccagct gacccagagc      480

cacaagtttc tgagcaccag cgtgggcgac cgggtgtcca tcacctgcaa agccagccag      540

gacgtgtaca acgccgtggc ctggtatcag cagaagcctg gccagagccc caagctgctg      600

atctacagcg ccagcagccg gtacaccggc gtgcccagca ggttcaccgg cagcggcagc      660

ggcccagact tcaccttcac catcagcagc gtgcaggccg aggacctggc cgtgtacttc      720

tgccagcagc acttccggac ccccttcacc ttcggctccg gcaccaagct ggaaatcaag      780

gccctgagca acagcatcat gtacttcagc cacttcgtgc ccgtgtttct gcccgccaag      840

cccaccacca cccctgcccc cagaccccct accccagccc ccacaatcgc cagccagccc      900

ctgagcctga ggcccgaggc cagcagacct gccgctgggg gagccgtgca caccaggggc      960

ctggacaagc ccttctgggt gctggtcgtg gtcggcggag tgctggcctg ttacagcctg     1020

ctggtcaccg tggccttcat catcttttgg gtccgcagca agcggagccg gctgctgcac     1080

agcgactaca tgaacatgac cccaaggcgg ccaggcccca cccggaagca ctaccagccc     1140

tatgcccctc ctagggactt cgccgcctac cggtccagag tgaagttcag ccgcagcgcc     1200

gacgcccctg cctaccagca gggccagaac cagctgtaca acgagctgaa cctgggcagg     1260

cgggaggaat acgacgtgct ggacaagcgc agaggccggg accctgagat gggcggcaag     1320

cccaggcgga agaaccccca ggaaggcctg tataacgaac tgcagaaaga caagatggcc     1380

gaggcctaca gcgagatcgg catgaagggc gagcggcgac gcggcaaggg ccacgacggc     1440

ctgtaccagg gcctgtccac cgccaccaag gacacctacg acgccctgca catgcaggcc     1500

ctgcctcccc gttaa                                                      1515


