                         SEQUENCE LISTING

<110>  The University of North Carolina at Chapel Hill
       The General Hospital Corporation
       Dotti, Gianpietro
       Ferrone, Soldano
       Hudson, Hannah Reid
       Dukhovlinova, Elena
       Ferrone, Cristina
       Wang, Xinhui
 
<120>  METHODS AND COMPOSITIONS FOR CHIMERIC ANTIGEN RECEPTOR TARGETING 
       CANCER CELLS

<130>  5470-837WO

<150>  US 62/684,513
<151>  2018-06-13

<160>  13    

<170>  PatentIn version 3.5

<210>  1
<211>  113
<212>  PRT
<213>  Artificial

<220>
<223>  UNC 763.74 Vh1

<400>  1

Gln Ile Gln Leu Val 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 Thr Phe Thr Asp Tyr 
            20                  25                  30          


Ser Met His Trp Val Lys Lys Thr Pro Gly Lys Gly Leu Lys Trp Leu 
        35                  40                  45              


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


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


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


Phe Ser Tyr Tyr Asp Tyr Trp Gly Gln Gly Thr Thr Leu Thr Val Ser 
            100                 105                 110         


Ser 
    


<210>  2
<211>  107
<212>  PRT
<213>  Artificial

<220>
<223>  UNC 763.74 Vk2

<400>  2

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


Glu Thr Val Ser Leu Ser Cys Arg Ala Ser Gln Thr Ile Tyr Lys Asn 
            20                  25                  30          


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


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


Ser Gly Ser Gly Thr Asp Tyr Thr Leu Asn Ile Asn Ser Val Lys Pro 
65                  70                  75                  80  


Glu Asp Glu Gly Ile Tyr Tyr Cys Leu Gln Gly Tyr Ser Thr Pro Trp 
                85                  90                  95      


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


<210>  3
<211>  122
<212>  PRT
<213>  Artificial

<220>
<223>  225.28 Vh1

<400>  3

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


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


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


Ala Glu Ile Arg Leu Lys Ser Asn Asn Phe Ala Arg Tyr Tyr Ala Glu 
    50                  55                  60                  


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


Val Tyr Leu Gln Met Ile Asn Leu Arg Ala Glu Asp Thr Gly Ile Tyr 
                85                  90                  95      


Tyr Cys Thr Ser Tyr Gly Asn Tyr Val Gly His Tyr Phe Asp His Trp 
            100                 105                 110         


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


<210>  4
<211>  107
<212>  PRT
<213>  Artificial

<220>
<223>  225.28 Vk

<400>  4

Asp Ile Val Met Thr Gln Ser Gln Lys Phe Met Ser Thr Ser Val Gly 
1               5                   10                  15      


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


Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Glu Pro Leu Leu 
        35                  40                  45              


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


Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Asn Val Gln Ser 
65                  70                  75                  80  


Glu Asp Leu Ala Glu Tyr Phe Cys Gln Gln Tyr Asn Ser Tyr Pro Leu 
                85                  90                  95      


Thr Phe Gly Gly Gly Thr Lys Leu Val Ile Lys 
            100                 105         


<210>  5
<211>  257
<212>  PRT
<213>  Artificial

<220>
<223>  UNC 763.74 scFv (Vk2.Vh1)

<400>  5

Ala Met Glu Phe Gly Leu Ser Trp Leu Phe Leu Val Ala Ile Leu Lys 
1               5                   10                  15      


Gly Val Gln Cys Asp Ile Leu Leu Thr Gln Ser Pro Ala Ile Leu Ser 
            20                  25                  30          


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


Ile Tyr Lys Asn Leu His Trp Tyr Gln Gln Lys Ser His Arg Ser Pro 
    50                  55                  60                  


Arg Leu Leu Ile Lys Tyr Gly Ser Asp Ser Ile Ser Gly Ile Pro Ser 
65                  70                  75                  80  


Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Tyr Thr Leu Asn Ile Asn 
                85                  90                  95      


Ser Val Lys Pro Glu Asp Glu Gly Ile Tyr Tyr Cys Leu Gln Gly Tyr 
            100                 105                 110         


Ser Thr Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Gly 
        115                 120                 125             


Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Gln Ile Gln 
    130                 135                 140                 


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


Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr Ser Met His 
                165                 170                 175     


Trp Val Lys Lys Thr Pro Gly Lys Gly Leu Lys Trp Leu Gly Trp Ile 
            180                 185                 190         


Asn Thr Ala Thr Gly Glu Pro Thr Tyr Ala Asp Asp Phe Lys Gly Arg 
        195                 200                 205             


Phe Ala Ile Ser Leu Glu Thr Ser Ala Arg Thr Val Tyr Leu Gln Ile 
    210                 215                 220                 


Asn Asn Leu Arg Asn Glu Asp Thr Ala Thr Tyr Phe Cys Phe Ser Tyr 
225                 230                 235                 240 


Tyr Asp Tyr Trp Gly Gln Gly Thr Thr Leu Thr Val Ser Ser Thr Arg 
                245                 250                 255     


Cys 
    


<210>  6
<211>  257
<212>  PRT
<213>  Artificial

<220>
<223>  UNC763.74 scFv (Vh1.Vk2)

<400>  6

Ala Met Glu Phe Gly Leu Ser Trp Leu Phe Leu Val Ala Ile Leu Lys 
1               5                   10                  15      


Gly Val Gln Cys Gln Ile Gln Leu Val Gln Ser Gly Pro Glu Leu Lys 
            20                  25                  30          


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


Phe Thr Asp Tyr Ser Met His Trp Val Lys Lys Thr Pro Gly Lys Gly 
    50                  55                  60                  


Leu Lys Trp Leu Gly Trp Ile Asn Thr Ala Thr Gly Glu Pro Thr Tyr 
65                  70                  75                  80  


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


Arg Thr Val Tyr Leu Gln Ile Asn Asn Leu Arg Asn Glu Asp Thr Ala 
            100                 105                 110         


Thr Tyr Phe Cys Phe Ser Tyr Tyr Asp Tyr Trp Gly Gln Gly Thr Thr 
        115                 120                 125             


Leu Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 
    130                 135                 140                 


Gly Gly Gly Asp Ile Leu Leu Thr Gln Ser Pro Ala Ile Leu Ser Val 
145                 150                 155                 160 


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


Tyr Lys Asn Leu His Trp Tyr Gln Gln Lys Ser His Arg Ser Pro Arg 
            180                 185                 190         


Leu Leu Ile Lys Tyr Gly Ser Asp Ser Ile Ser Gly Ile Pro Ser Arg 
        195                 200                 205             


Phe Thr Gly Ser Gly Ser Gly Thr Gly Tyr Thr Leu Asn Ile Asn Ser 
    210                 215                 220                 


Val Lys Pro Glu Asp Glu Gly Ile Tyr Tyr Cys Leu Gln Gly Tyr Ser 
225                 230                 235                 240 


Thr Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Thr Arg 
                245                 250                 255     


Cys 
    


<210>  7
<211>  266
<212>  PRT
<213>  Artificial

<220>
<223>  225.28 scFv (Vh1.Vk)

<400>  7

Ala Met Glu Phe Gly Leu Ser Trp Leu Phe Leu Val Ala Ile Leu Lys 
1               5                   10                  15      


Gly Val Gln Cys Lys Val Lys Leu Gln Glu Ser Gly Gly Gly Leu Val 
            20                  25                  30          


Gln Pro Gly Gly Ser Met Lys Leu Ser Cys Val Val Ser Gly Phe Thr 
        35                  40                  45              


Phe Ser Asn Tyr Trp Met Asn Trp Val Arg Gln Ser Pro Glu Lys Gly 
    50                  55                  60                  


Leu Glu Trp Ile Ala Glu Ile Arg Leu Lys Ser Asn Asn Phe Ala Arg 
65                  70                  75                  80  


Tyr Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp 
                85                  90                  95      


Ser Lys Ser Ser Val Tyr Leu Gln Met Ile Asn Leu Arg Ala Glu Asp 
            100                 105                 110         


Thr Gly Ile Tyr Tyr Cys Thr Ser Tyr Gly Asn Tyr Val Gly His Tyr 
        115                 120                 125             


Phe Asp His Trp Gly Gln Gly Thr Thr Leu Thr Val Ser Ser Gly Gly 
    130                 135                 140                 


Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Asp Ile Val Met 
145                 150                 155                 160 


Thr Gln Ser Gln Lys Phe Met Ser Thr Ser Val Gly Asp Arg Val Ser 
                165                 170                 175     


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


Gln Gln Lys Pro Gly Gln Ser Pro Glu Pro Leu Leu Phe Ser Ala Ser 
        195                 200                 205             


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


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


Glu Tyr Phe Cys Gln Gln Tyr Asn Ser Tyr Pro Leu Thr Phe Gly Gly 
                245                 250                 255     


Gly Thr Lys Leu Val Ile Lys Thr Arg Cys 
            260                 265     


<210>  8
<211>  266
<212>  PRT
<213>  Artificial

<220>
<223>  225.28 scFv (Vk.Vh1)

<400>  8

Ala Met Glu Phe Gly Leu Ser Trp Leu Phe Leu Val Ala Ile Leu Lys 
1               5                   10                  15      


Gly Val Gln Cys Asp Ile Val Met Thr Gln Ser Gln Lys Phe Met Ser 
            20                  25                  30          


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


Val Asp Thr Asn Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro 
    50                  55                  60                  


Glu Pro Leu Leu Phe Ser Ala Ser Tyr Arg Tyr Thr Gly Val Pro Asp 
65                  70                  75                  80  


Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser 
                85                  90                  95      


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


Ser Tyr Pro Leu Thr Phe Gly Gly Gly Thr Lys Leu Val Ile Lys Gly 
        115                 120                 125             


Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Lys Val Lys 
    130                 135                 140                 


Leu Gln Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Met Lys 
145                 150                 155                 160 


Leu Ser Cys Val Val Ser Gly Phe Thr Phe Ser Asn Tyr Trp Met Asn 
                165                 170                 175     


Trp Val Arg Gln Ser Pro Glu Lys Gly Leu Glu Trp Ile Ala Glu Ile 
            180                 185                 190         


Arg Leu Lys Ser Asn Asn Phe Ala Arg Tyr Tyr Ala Glu Ser Val Lys 
        195                 200                 205             


Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Ser Ser Val Tyr Leu 
    210                 215                 220                 


Gln Met Ile Asn Leu Arg Ala Glu Asp Thr Gly Ile Tyr Tyr Cys Thr 
225                 230                 235                 240 


Ser Tyr Gly Asn Tyr Val Gly His Tyr Phe Asp His Trp Gly Gln Gly 
                245                 250                 255     


Thr Thr Leu Thr Val Ser Ser Thr Arg Cys 
            260                 265     


<210>  9
<211>  773
<212>  DNA
<213>  Artificial

<220>
<223>  scFv.763.74 VK2.VH1

<400>  9
tagccatgga attcggcctg agctggctgt tcctggtggc catcctgaag ggcgtgcagt       60

gcgatatcct gctcacccag tctccagcaa tcctgtctgt gactccagga gaaacagtca      120

gtctttcctg tagggccagc cagactattt acaagaacct acactggtat caacagaaat      180

cacatcggtc tccaaggctt ctcatcaagt atggttctga ttccatctct gggatcccct      240

ccaggttcac tggcagtgga tcagggacag attacactct caatatcaac agtgtgaagc      300

ccgaagatga aggaatatat tactgtcttc aaggttacag tacaccttgg acgttcggtg      360

gaggcaccaa gctggaaatc aaaggtgggg gcggttcagg cggaggtggc tctggcggtg      420

gcggacagat ccagttggtg cagtctggac ctgagctgaa gaagcctgga gagacagtca      480

agatctcctg caaggcttct ggttatacct tcacagacta ttcaatgcac tgggtgaaga      540

agactccagg aaagggttta aagtggctgg gctggataaa cactgcgact ggtgagccaa      600

catatgcaga tgacttcaag ggacggtttg ccatctcttt ggaaacctct gccaggactg      660

tctatttgca gatcaataat ctcagaaatg aggacacggc tacatatttc tgttttagtt      720

actacgacta ctggggccaa ggcaccactc tcacagtttc ctcaacgcgt tgc             773


<210>  10
<211>  773
<212>  DNA
<213>  Artificial

<220>
<223>  scFv.763.74 VH1.VK2

<400>  10
tagccatgga attcggcctg agctggctgt tcctggtggc catcctgaag ggcgtgcagt       60

gccagatcca gttggtgcag tctggacctg agctgaagaa gcctggagag acagtcaaga      120

tctcctgcaa ggcttctggt tataccttca cagactattc aatgcactgg gtgaagaaga      180

ctccaggaaa gggtttaaag tggctgggct ggataaacac tgcgactggt gagccaacat      240

atgcagatga cttcaaggga cggtttgcca tctctttgga aacctctgcc aggactgtct      300

atttgcagat caataatctc agaaatgagg acacggctac atatttctgt tttagttact      360

acgactactg gggccaaggc accactctca cagtttcctc aggtgggggc ggttcaggcg      420

gaggtggctc tggcggtggc ggagatatcc tgctcaccca gtctccagca atcctgtctg      480

tgactccagg agaaacagtc agtctttcct gtagggccag ccagactatt tacaagaacc      540

tacactggta tcaacagaaa tcacatcggt ctccaaggct tctcatcaag tatggttctg      600

attccatctc tgggatcccc tccaggttca ctggcagtgg atcagggaca gattacactc      660

tcaatatcaa cagtgtgaag cccgaagatg aaggaatata ttactgtctt caaggttaca      720

gtacaccttg gacgttcggt ggaggcacca agctggaaat caaaacgcgt tgc             773


<210>  11
<211>  800
<212>  DNA
<213>  Artificial

<220>
<223>  scFv225.28 VH1.VK

<400>  11
tagccatgga attcggcctg agctggctgt tcctggtggc catcctgaag ggcgtgcagt       60

gcaaggtgaa gctgcaggag tctggaggag gcttggtgca acctggagga tccatgaaac      120

tctcctgtgt tgtctctgga ttcactttca gtaattactg gatgaactgg gtccgccagt      180

ctccagagaa ggggcttgag tggattgcag aaattagatt gaaatccaat aattttgcaa      240

gatattatgc ggagtctgtg aaagggaggt tcaccatctc aagagatgat tccaaaagta      300

gtgtctacct gcaaatgatc aacctaagag ctgaagatac tggcatttat tactgtacca      360

gttatggtaa ctacgttggg cactattttg accactgggg ccaaggcacc actctcacag      420

tctcctcagg tgggggcggt tcaggcggag gtggctctgg cggtggcgga gacattgtga      480

tgacccagtc tcaaaaattc atgtccacat cagtaggaga cagggtcagc gtcacctgca      540

aggccagtca gaatgtggat actaatgtag cctggtatca acaaaaacca gggcaatctc      600

ctgaaccact gcttttctcg gcatcctacc gttacactgg agtccctgat cgcttcacag      660

gcagtggatc tgggacagat ttcactctca ccatcagcaa tgtgcagtct gaagacttgg      720

cagagtattt ctgtcagcaa tataacagct atcctctgac gttcggtgga ggcaccaagc      780

tggtgatcaa aacgcgttgc                                                  800


<210>  12
<211>  800
<212>  DNA
<213>  Artificial

<220>
<223>  scFv225.28 VK.VH1

<400>  12
tagccatgga attcggcctg agctggctgt tcctggtggc catcctgaag ggcgtgcagt       60

gcgacattgt gatgacccag tctcaaaaat tcatgtccac atcagtagga gacagggtca      120

gcgtcacctg caaggccagt cagaatgtgg atactaatgt agcctggtat caacaaaaac      180

cagggcaatc tcctgaacca ctgcttttct cggcatccta ccgttacact ggagtccctg      240

atcgcttcac aggcagtgga tctgggacag atttcactct caccatcagc aatgtgcagt      300

ctgaagactt ggcagagtat ttctgtcagc aatataacag ctatcctctg acgttcggtg      360

gaggcaccaa gctggtgatc aaaggtgggg gcggttcagg cggaggtggc tctggcggtg      420

gcggaaaggt gaagctgcag gagtctggag gaggcttggt gcaacctgga ggatccatga      480

aactctcctg tgttgtctct ggattcactt tcagtaatta ctggatgaac tgggtccgcc      540

agtctccaga gaaggggctt gagtggattg cagaaattag attgaaatcc aataattttg      600

caagatatta tgcggagtct gtgaaaggga ggttcaccat ctcaagagat gattccaaaa      660

gtagtgtcta cctgcaaatg atcaacctaa gagctgaaga tactggcatt tattactgta      720

ccagttatgg taactacgtt gggcactatt ttgaccactg gggccaaggc accactctca      780

cagtctcctc aacgcgttgc                                                  800


<210>  13
<211>  15
<212>  PRT
<213>  Artificial

<220>
<223>  linker peptide

<400>  13

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


