
                                SEQUENCE LISTING

<110> Michael C. Jensen
      James F. Matthaei

<120> METHODS AND COMPOSITIONS FOR STIMULATION
  OF CHIMERIC ANTIGEN RECEPTOR T CELLS WITH HAPTEN LABELLED
  CELLS

<130> SCRI.272WO

<150> 62969917            
<151> 2020-02-04  

<160> 21

<170> FastSEQ for Windows Version 4.0

<210> 1
<211> 251
<212> PRT
<213> Artifiicial Sequence

<220> 
<223> FITCE2 scFv

<400> 1
Ser Val Leu Thr Gln Pro Ser Ser Val Ser Ala Ala Pro Gly Gln Lys
 1               5                  10                  15      
Val Thr Ile Ser Cys Ser Gly Ser Thr Ser Asn Ile Gly Asn Asn Tyr
            20                  25                  30          
Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu Met Ile
        35                  40                  45              
Tyr Asp Val Ser Lys Arg Pro Ser Gly Val Pro Asp Arg Phe Ser Gly
    50                  55                  60                  
Ser Lys Ser Gly Asn Ser Ala Ser Leu Asp Ile Ser Gly Leu Gln Ser
65                  70                  75                  80  
Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu Ser
                85                  90                  95      
Glu Phe Leu Phe Gly Thr Gly Thr Lys Leu Thr Val Leu Gly Gly Gly
            100                 105                 110         
Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln
        115                 120                 125             
Leu Val Glu Ser Gly Gly Asn Leu Val Gln Pro Gly Gly Ser Leu Arg
    130                 135                 140                 
Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Gly Ser Phe Ser Met Ser
145                 150                 155                 160 
Trp Val Arg Gln Ala Pro Gly Gly Gly Leu Glu Trp Val Ala Gly Leu
                165                 170                 175     
Ser Ala Arg Ser Ser Leu Thr His Tyr Ala Asp Ser Val Lys Gly Arg
            180                 185                 190         
Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Val Tyr Leu Gln Met
        195                 200                 205             
Asn Ser Leu Arg Val Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Arg
    210                 215                 220                 
Ser Tyr Asp Ser Ser Gly Tyr Trp Gly His Phe Tyr Ser Tyr Met Asp
225                 230                 235                 240 
Val Trp Gly Gln Gly Thr Leu Val Thr Val Ser
                245                 250     


<210> 2
<211> 251
<212> PRT
<213> Artificial Sequence

<220> 
<223> FITCE2 TyrH133Ala scFv

<400> 2
Ser Val Leu Thr Gln Pro Ser Ser Val Ser Ala Ala Pro Gly Gln Lys
 1               5                  10                  15      
Val Thr Ile Ser Cys Ser Gly Ser Thr Ser Asn Ile Gly Asn Asn Tyr
            20                  25                  30          
Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu Met Ile
        35                  40                  45              
Tyr Asp Val Ser Lys Arg Pro Ser Gly Val Pro Asp Arg Phe Ser Gly
    50                  55                  60                  
Ser Lys Ser Gly Asn Ser Ala Ser Leu Asp Ile Ser Gly Leu Gln Ser
65                  70                  75                  80  
Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu Ser
                85                  90                  95      
Glu Phe Leu Phe Gly Thr Gly Thr Lys Leu Thr Val Leu Gly Gly Gly
            100                 105                 110         
Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln
        115                 120                 125             
Leu Val Glu Ser Gly Gly Asn Leu Val Gln Pro Gly Gly Ser Leu Arg
    130                 135                 140                 
Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Gly Ser Phe Ser Met Ser
145                 150                 155                 160 
Trp Val Arg Gln Ala Pro Gly Gly Gly Leu Glu Trp Val Ala Gly Leu
                165                 170                 175     
Ser Ala Arg Ser Ser Leu Thr His Tyr Ala Asp Ser Val Lys Gly Arg
            180                 185                 190         
Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Val Tyr Leu Gln Met
        195                 200                 205             
Asn Ser Leu Arg Val Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Arg
    210                 215                 220                 
Ser Tyr Asp Ser Ser Gly Tyr Trp Gly His Phe Ala Ser Tyr Met Asp
225                 230                 235                 240 
Val Trp Gly Gln Gly Thr Leu Val Thr Val Ser
                245                 250     


<210> 3
<211> 250
<212> PRT
<213> Artificial Sequence

<220> 
<223> FITCE2 HisH131Ala scFv

<400> 3
Val Leu Thr Gln Pro Ser Ser Val Ser Ala Ala Pro Gly Gln Lys Val
 1               5                  10                  15      
Thr Ile Ser Cys Ser Gly Ser Thr Ser Asn Ile Gly Asn Asn Tyr Val
            20                  25                  30          
Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu Met Ile Tyr
        35                  40                  45              
Asp Val Ser Lys Arg Pro Ser Gly Val Pro Asp Arg Phe Ser Gly Ser
    50                  55                  60                  
Lys Ser Gly Asn Ser Ala Ser Leu Asp Ile Ser Gly Leu Gln Ser Glu
65                  70                  75                  80  
Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu Ser Glu
                85                  90                  95      
Phe Leu Phe Gly Thr Gly Thr Lys Leu Thr Val Leu Gly Gly Gly Gly
            100                 105                 110         
Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu
        115                 120                 125             
Val Glu Ser Gly Gly Asn Leu Val Gln Pro Gly Gly Ser Leu Arg Leu
    130                 135                 140                 
Ser Cys Ala Ala Ser Gly Phe Thr Phe Gly Ser Phe Ser Met Ser Trp
145                 150                 155                 160 
Val Arg Gln Ala Pro Gly Gly Gly Leu Glu Trp Val Ala Gly Leu Ser
                165                 170                 175     
Ala Arg Ser Ser Leu Thr His Tyr Ala Asp Ser Val Lys Gly Arg Phe
            180                 185                 190         
Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Val Tyr Leu Gln Met Asn
        195                 200                 205             
Ser Leu Arg Val Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Arg Ser
    210                 215                 220                 
Tyr Asp Ser Ser Gly Tyr Trp Gly Ala Phe Tyr Ser Tyr Met Asp Val
225                 230                 235                 240 
Trp Gly Gln Gly Thr Leu Val Thr Val Ser
                245                 250 


<210> 4
<211> 254
<212> PRT
<213> Artificial Sequence

<220> 
<223> 4M5.3 scFv

<400> 4
Asp Val Val Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly
 1               5                  10                  15      
Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Val His Ser
            20                  25                  30          
Asn Gly Asn Thr Tyr Leu Arg Trp Tyr Leu Gln Lys Pro Gly Gln Ser
        35                  40                  45              
Pro Lys Val Leu Ile Tyr Lys Val Ser Asn Arg Val Ser Gly Val Pro
    50                  55                  60                  
Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile
65                  70                  75                  80  
Asn Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Phe Cys Ser Gln Ser
                85                  90                  95      
Thr His Val Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys
            100                 105                 110         
Ser Ser Ala Asp Asp Ala Lys Lys Asp Ala Ala Lys Lys Asp Asp Ala
        115                 120                 125             
Lys Lys Asp Asp Ala Lys Lys Asp Gly Gly Val Lys Leu Asp Glu Thr
    130                 135                 140                 
Gly Gly Gly Leu Val Gln Pro Gly Gly Ala Met Lys Leu Ser Cys Val
145                 150                 155                 160 
Thr Ser Gly Phe Thr Phe Gly His Tyr Trp Met Asn Trp Val Arg Gln
                165                 170                 175     
Ser Pro Glu Lys Gly Leu Glu Trp Val Ala Gln Phe Arg Asn Lys Pro
            180                 185                 190         
Tyr Asn Tyr Glu Thr Tyr Tyr Ser Asp Ser Val Lys Gly Arg Phe Thr
        195                 200                 205             
Ile Ser Arg Asp Asp Ser Lys Ser Ser Val Tyr Leu Gln Met Asn Asn
    210                 215                 220                 
Leu Arg Val Glu Asp Thr Gly Ile Tyr Tyr Cys Thr Gly Ala Ser Tyr
225                 230                 235                 240 
Gly Met Glu Tyr Leu Gly Gln Gly Thr Ser Val Thr Val Ser
                245                 250                 


<210> 5
<211> 265
<212> PRT
<213> Artificial Sequence

<220> 
<223> 4D5Flu scFv

<400> 5
Asp Tyr Lys Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala
 1               5                  10                  15      
Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Leu
            20                  25                  30          
Val His Ser Gln Gly Asn Thr Tyr Leu Arg Trp Tyr Gln Gln Lys Pro
        35                  40                  45              
Gly Lys Ala Pro Lys Val Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser
    50                  55                  60                  
Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr
65                  70                  75                  80  
Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys
                85                  90                  95      
Gln Gln Ser Thr His Val Pro Trp Thr Phe Gly Gln Gly Thr Lys Val
            100                 105                 110         
Glu Leu Lys Arg Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
        115                 120                 125             
Gly Gly Gly Ser Ser Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly
    130                 135                 140                 
Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln
145                 150                 155                 160 
Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe
                165                 170                 175     
Ser Asp Tyr Trp Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu
            180                 185                 190         
Glu Trp Val Ala Gln Ile Arg Asn Lys Pro Tyr Asn Tyr Glu Thr Tyr
        195                 200                 205             
Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Thr Ser
    210                 215                 220                 
Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr
225                 230                 235                 240 
Ala Val Tyr Tyr Cys Thr Gly Ser Tyr Tyr Gly Met Asp Tyr Trp Gly
                245                 250                 255     
Gln Gly Thr Leu Val Thr Val Ser Ser
            260                 265 


<210> 6
<211> 244
<212> PRT
<213> Artificial Sequence

<220> 
<223> 4420

<400> 6
Asp Val Val Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly
 1               5                  10                  15      
Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Val His Ser
            20                  25                  30          
Gln Gly Asn Thr Tyr Leu Arg Trp Tyr Leu Gln Lys Pro Gly Gln Ser
        35                  40                  45              
Pro Lys Val Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro
    50                  55                  60                  
Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile
65                  70                  75                  80  
Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Phe Cys Ser Gln Ser
                85                  90                  95      
Thr His Val Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Gly
            100                 105                 110         
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val
        115                 120                 125             
Lys Leu Asp Glu Thr Gly Gly Gly Leu Val Gln Pro Gly Arg Pro Met
    130                 135                 140                 
Lys Leu Ser Cys Val Ala Ser Gly Phe Thr Phe Ser Asp Tyr Trp Met
145                 150                 155                 160 
Asn Trp Val Arg Gln Ser Pro Glu Lys Gly Leu Glu Trp Val Ala Gln
                165                 170                 175     
Ile Arg Asn Lys Pro Tyr Asn Tyr Glu Thr Tyr Tyr Ser Asp Ser Val
            180                 185                 190         
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Ser Ser Val Tyr
        195                 200                 205             
Leu Gln Met Asn Asn Leu Arg Val Glu Asp Met Gly Ile Tyr Tyr Cys
    210                 215                 220                 
Thr Gly Ser Tyr Tyr Gly Met Asp Tyr Trp Gly Gln Gly Thr Ser Val
225                 230                 235                 240 
Thr Val Ser Ser
                


<210> 7
<211> 789
<212> DNA
<213> Artificial Sequence

<220> 
<223> anti DNP scFv
      (VH to VL from rabbit
      

<400> 7
cagtgtcagc agctggagca gtccggagga ggagccgaag gaggcctggt caagcctggg 60
ggatccctgg aactctgctg caaagcctct ggattctccc tcagtagtag ctactgcata 120
tgttgggtcc gccaggctcc agggaagggg ctggagtgga tcggatgcat ttatgctggt 180
agtagtggta gcacttacta cgcgagctgg gtgaatggcc gattcactct ctccagagac 240
attgaccaga gcacaggttg cctacaactg aacagtctga cagccgcgga cacggccatg 300
tattactgtg cgagagcccc ctatagtagt ggctgggtcc tctactttaa cttgtggggc 360
ccaggcaccc tggtcattgt ctcctcaggc ggagggggct ctggcggcgg aggatctggg 420
ggagggggca gcccaggtgc cacatttgcc caagtgctga cccagactcc atcgcctgtg 480
tctgcagctg tgggaggcac agtcaccatc agttgccagt ccagtgagag tgtttatggt 540
aacagccgct tagcctggta tcagcagaaa ccagggcagt ctcccaagct cctgatctat 600
tatgcatcca ctctggcatc tggggtccct tcgcggttca aaggcagtgg atctgggaca 660
cagttcactc tcaccattag cgacctggag tgtgacgatg ctgcctctta ctactgtcaa 720
ggcggttatt atagtggtaa tcttgatgcg cttgctttcg gcggagggac cgaggtggtg 780
gtcagaggt                                                         789

<210> 8
<211> 161
<212> PRT
<213> Artificial Sequence

<220> 
<223> anti DNP scFv
      (VH to VL from rabbit)

<400> 8
Cys Ala Arg Ala Pro Tyr Ser Ser Gly Trp Val Leu Tyr Phe Asn Leu
 1               5                  10                  15      
Trp Gly Pro Gly Thr Leu Val Ile Val Ser Ser Gly Gly Gly Gly Ser
            20                  25                  30          
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Pro Gly Ala Thr Phe Ala
        35                  40                  45              
Gln Val Leu Thr Gln Thr Pro Ser Pro Val Ser Ala Ala Val Gly Gly
    50                  55                  60                  
Thr Val Thr Ile Ser Cys Gln Ser Ser Glu Ser Val Tyr Gly Asn Ser
65                  70                  75                  80  
Arg Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu
                85                  90                  95      
Ile Tyr Tyr Ala Ser Thr Leu Ala Ser Gly Val Pro Ser Arg Phe Lys
            100                 105                 110         
Gly Ser Gly Ser Gly Thr Gln Phe Thr Leu Thr Ile Ser Asp Leu Glu
        115                 120                 125             
Cys Asp Asp Ala Ala Ser Tyr Tyr Cys Gln Gly Gly Tyr Tyr Ser Gly
    130                 135                 140                 
Asn Leu Asp Ala Leu Ala Phe Gly Gly Gly Thr Glu Val Val Val Arg
145                 150                 155                 160 
Gly
    


<210> 9
<211> 789
<212> DNA
<213> Artificial Sequence

<220> 
<223> anti DNP scFv
      (VL to VH from rabbit)
      

<400> 9
ccaggtgcca catttgccca agtgctgacc cagactccat cgcctgtgtc tgcagctgtg 60
ggaggcacag tcaccatcag ttgccagtcc agtgagagtg tttatggtaa cagccgctta 120
gcctggtatc agcagaaacc agggcagtct cccaagctcc tgatctatta tgcatccact 180
ctggcatctg gggtcccttc gcggttcaaa ggcagtggat ctgggacaca gttcactctc 240
accattagcg acctggagtg tgacgatgct gcctcttact actgtcaagg cggttattat 300
agtggtaatc ttgatgcgct tgctttcggc ggagggaccg aggtggtggt cagaggtggc 360
ggagggggct ctggcggcgg aggatctggg ggagggggca gccagtgtca gcagctggag 420
cagtccggag gaggagccga aggaggcctg gtcaagcctg ggggatccct ggaactctgc 480
tgcaaagcct ctggattctc cctcagtagt agctactgca tatgttgggt ccgccaggct 540
ccagggaagg ggctggagtg gatcggatgc atttatgctg gtagtagtgg tagcacttac 600
tacgcgagct gggtgaatgg ccgattcact ctctccagag acattgacca gagcacaggt 660
tgcctacaac tgaacagtct gacagccgcg gacacggcca tgtattactg tgcgagagcc 720
ccctatagta gtggctgggt cctctacttt aacttgtggg gcccaggcac cctggtcatt 780
gtctcctca                                                         789

<210> 10
<211> 263
<212> PRT
<213> Artificial Sequence

<220> 
<223> anti DNP scFv
      (VL to VH from rabbit)
      

<400> 10
Pro Gly Ala Thr Phe Ala Gln Val Leu Thr Gln Thr Pro Ser Pro Val
 1               5                  10                  15      
Ser Ala Ala Val Gly Gly Thr Val Thr Ile Ser Cys Gln Ser Ser Glu
            20                  25                  30          
Ser Val Tyr Gly Asn Ser Arg Leu Ala Trp Tyr Gln Gln Lys Pro Gly
        35                  40                  45              
Gln Ser Pro Lys Leu Leu Ile Tyr Tyr Ala Ser Thr Leu Ala Ser Gly
    50                  55                  60                  
Val Pro Ser Arg Phe Lys Gly Ser Gly Ser Gly Thr Gln Phe Thr Leu
65                  70                  75                  80  
Thr Ile Ser Asp Leu Glu Cys Asp Asp Ala Ala Ser Tyr Tyr Cys Gln
                85                  90                  95      
Gly Gly Tyr Tyr Ser Gly Asn Leu Asp Ala Leu Ala Phe Gly Gly Gly
            100                 105                 110         
Thr Glu Val Val Val Arg Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly
        115                 120                 125             
Ser Gly Gly Gly Gly Ser Gln Cys Gln Gln Leu Glu Gln Ser Gly Gly
    130                 135                 140                 
Gly Ala Glu Gly Gly Leu Val Lys Pro Gly Gly Ser Leu Glu Leu Cys
145                 150                 155                 160 
Cys Lys Ala Ser Gly Phe Ser Leu Ser Ser Ser Tyr Cys Ile Cys Trp
                165                 170                 175     
Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile Gly Cys Ile Tyr
            180                 185                 190         
Ala Gly Ser Ser Gly Ser Thr Tyr Tyr Ala Ser Trp Val Asn Gly Arg
        195                 200                 205             
Phe Thr Leu Ser Arg Asp Ile Asp Gln Ser Thr Gly Cys Leu Gln Leu
    210                 215                 220                 
Asn Ser Leu Thr Ala Ala Asp Thr Ala Met Tyr Tyr Cys Ala Arg Ala
225                 230                 235                 240 
Pro Tyr Ser Ser Gly Trp Val Leu Tyr Phe Asn Leu Trp Gly Pro Gly
                245                 250                 255     
Thr Leu Val Ile Val Ser Ser
            260             


<210> 11
<211> 22
<212> PRT
<213> Artificial Sequence

<220> 
<223> GM-CSF

<400> 11
Met Leu Leu Leu Val Thr Ser Leu Leu Leu Cys Glu Leu Pro His Pro
 1               5                  10                  15      
Ala Phe Leu Leu Ile Pro
            20          


<210> 12
<211> 229
<212> PRT
<213> Artificial Sequence

<220> 
<223> Spacer (long): IgG4hinge-CH2(L235D)-CH3

<400> 12
Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe
 1               5                  10                  15      
Asp Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr
            20                  25                  30          
Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val
        35                  40                  45              
Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val
    50                  55                  60                  
Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser
65                  70                  75                  80  
Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu
                85                  90                  95      
Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser
            100                 105                 110         
Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro
        115                 120                 125             
Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln
    130                 135                 140                 
Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala
145                 150                 155                 160 
Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr
                165                 170                 175     
Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu
            180                 185                 190         
Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser
        195                 200                 205             
Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser
    210                 215                 220                 
Leu Ser Leu Gly Lys
225                 


<210> 13
<211> 119
<212> PRT
<213> Artificial Sequence

<220> 
<223> Spacer (medium): IgG4 hinge-CH3

<400> 13
Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Gly Gln Pro Arg
 1               5                  10                  15      
Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys
            20                  25                  30          
Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp
        35                  40                  45              
Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys
    50                  55                  60                  
Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser
65                  70                  75                  80  
Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser
                85                  90                  95      
Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser
            100                 105                 110         
Leu Ser Leu Ser Leu Gly Lys
        115                 


<210> 14
<211> 12
<212> PRT
<213> Artificial Sequence

<220> 
<223> Spacer (short): IgG4 hinged

<400> 14
Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro
 1               5                  10          


<210> 15
<211> 28
<212> PRT
<213> Artificial Sequence

<220> 
<223> CD28tm

<400> 15
Met Phe Trp Val Leu Val Val Val Gly Gly Val Leu Ala Cys Tyr Ser
 1               5                  10                  15      
Leu Leu Val Thr Val Ala Phe Ile Ile Phe Trp Val
            20                  25              


<210> 16
<211> 42
<212> PRT
<213> Artificial Sequence

<220> 
<223> 4-1BB

<400> 16
Lys Arg Gly Arg Lys Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met
 1               5                  10                  15      
Arg Pro Val Gln Thr Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe
            20                  25                  30          
Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu
        35                  40          


<210> 17
<211> 112
<212> PRT
<213> Artificial Sequence

<220> 
<223> CD3 zeta

<400> 17
Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly
 1               5                  10                  15      
Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr
            20                  25                  30          
Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys
        35                  40                  45              
Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys
    50                  55                  60                  
Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg
65                  70                  75                  80  
Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala
                85                  90                  95      
Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg
            100                 105                 110         


<210> 18
<211> 21
<212> PRT
<213> Artificial Sequence

<220> 
<223> T2A

<400> 18
Gly Gly Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly Asp Val Glu
 1               5                  10                  15      
Glu Asn Pro Gly Pro
            20      


<210> 19
<211> 357
<212> PRT
<213> Artificial Sequence

<220> 
<223> GM-CSF receptor ss to EGFRt

<400> 19
Met Leu Leu Leu Val Thr Ser Leu Leu Leu Cys Glu Leu Pro His Pro
 1               5                  10                  15      
Ala Phe Leu Leu Ile Pro Arg Lys Val Cys Asn Gly Ile Gly Ile Gly
            20                  25                  30          
Glu Phe Lys Asp Ser Leu Ser Ile Asn Ala Thr Asn Ile Lys His Phe
        35                  40                  45              
Lys Asn Cys Thr Ser Ile Ser Gly Asp Leu His Ile Leu Pro Val Ala
    50                  55                  60                  
Phe Arg Gly Asp Ser Phe Thr His Thr Pro Pro Leu Asp Pro Gln Glu
65                  70                  75                  80  
Leu Asp Ile Leu Lys Thr Val Lys Glu Ile Thr Gly Phe Leu Leu Ile
                85                  90                  95      
Gln Ala Trp Pro Glu Asn Arg Thr Asp Leu His Ala Phe Glu Asn Leu
            100                 105                 110         
Glu Ile Ile Arg Gly Arg Thr Lys Gln His Gly Gln Phe Ser Leu Ala
        115                 120                 125             
Val Val Ser Leu Asn Ile Thr Ser Leu Gly Leu Arg Ser Leu Lys Glu
    130                 135                 140                 
Ile Ser Asp Gly Asp Val Ile Ile Ser Gly Asn Lys Asn Leu Cys Tyr
145                 150                 155                 160 
Ala Asn Thr Ile Asn Trp Lys Lys Leu Phe Gly Thr Ser Gly Gln Lys
                165                 170                 175     
Thr Lys Ile Ile Ser Asn Arg Gly Glu Asn Ser Cys Lys Ala Thr Gly
            180                 185                 190         
Gln Val Cys His Ala Leu Cys Ser Pro Glu Gly Cys Trp Gly Pro Glu
        195                 200                 205             
Pro Arg Asp Cys Val Ser Cys Arg Asn Val Ser Arg Gly Arg Glu Cys
    210                 215                 220                 
Val Asp Lys Cys Asn Leu Leu Glu Gly Glu Pro Arg Glu Phe Val Glu
225                 230                 235                 240 
Asn Ser Glu Cys Ile Gln Cys His Pro Glu Cys Leu Pro Gln Ala Met
                245                 250                 255     
Asn Ile Thr Cys Thr Gly Arg Gly Pro Asp Asn Cys Ile Gln Cys Ala
            260                 265                 270         
His Tyr Ile Asp Gly Pro His Cys Val Lys Thr Cys Pro Ala Gly Val
        275                 280                 285             
Met Gly Glu Asn Asn Thr Leu Val Trp Lys Tyr Ala Asp Ala Gly His
    290                 295                 300                 
Val Cys His Leu Cys His Pro Asn Cys Thr Tyr Gly Cys Thr Gly Pro
305                 310                 315                 320 
Gly Leu Glu Gly Cys Pro Thr Asn Gly Pro Lys Ile Pro Ser Ile Ala
                325                 330                 335     
Thr Gly Met Val Gly Ala Leu Leu Leu Leu Leu Val Val Ala Leu Gly
            340                 345                 350         
Ile Gly Leu Phe Met
        355         


<210> 20
<211> 187
<212> PRT
<213> Artificial Sequence

<220> 
<223> DHFRdm

<400> 20
Met Val Gly Ser Leu Asn Cys Ile Val Ala Val Ser Gln Asn Met Gly
 1               5                  10                  15      
Ile Gly Lys Asn Gly Asp Phe Pro Trp Pro Pro Leu Arg Asn Glu Ser
            20                  25                  30          
Arg Tyr Phe Gln Arg Met Thr Thr Thr Ser Ser Val Glu Gly Lys Gln
        35                  40                  45              
Asn Leu Val Ile Met Gly Lys Lys Thr Trp Phe Ser Ile Pro Glu Lys
    50                  55                  60                  
Asn Arg Pro Leu Lys Gly Arg Ile Asn Leu Val Leu Ser Arg Glu Leu
65                  70                  75                  80  
Lys Glu Pro Pro Gln Gly Ala His Phe Leu Ser Arg Ser Leu Asp Asp
                85                  90                  95      
Ala Leu Lys Leu Thr Glu Gln Pro Glu Leu Ala Asn Lys Val Asp Met
            100                 105                 110         
Val Trp Ile Val Gly Gly Ser Ser Val Tyr Lys Glu Ala Met Asn His
        115                 120                 125             
Pro Gly His Leu Lys Leu Phe Val Thr Arg Ile Met Gln Asp Phe Glu
    130                 135                 140                 
Ser Asp Thr Phe Phe Pro Glu Ile Asp Leu Glu Lys Tyr Lys Leu Leu
145                 150                 155                 160 
Pro Glu Tyr Pro Gly Val Leu Ser Asp Val Gln Glu Glu Lys Gly Ile
                165                 170                 175     
Lys Tyr Lys Phe Glu Val Tyr Glu Lys Asn Asp
            180                 185         


<210> 21
<211> 1751
<212> PRT
<213> Artificial Sequence

<220> 
<223> Dual CAR sequence: mAb806 VHVL
      scFv-IgG4hinge-CD28tm/CD28gg-Zeta-T2A-EGFRt-P2-Ant
      iFL(FITC-E2 Tyr100gAla)scFv-IgG4hinge-CH2(L235D,
      N297Q)-CH3--CD28tm/41BB-zeta-T2A-DHFRdm-epHIV7.2

<400> 21
Met Leu Leu Leu Val Thr Ser Leu Leu Leu Cys Glu Leu Pro His Pro
 1               5                  10                  15      
Ala Phe Leu Leu Ile Pro Asp Val Gln Leu Gln Glu Ser Gly Pro Ser
            20                  25                  30          
Leu Val Lys Pro Ser Gln Ser Leu Ser Leu Thr Cys Thr Val Thr Gly
        35                  40                  45              
Tyr Ser Ile Thr Ser Asp Phe Ala Trp Asn Trp Ile Arg Gln Phe Pro
    50                  55                  60                  
Gly Asn Lys Leu Glu Trp Met Gly Tyr Ile Ser Tyr Ser Gly Asn Thr
65                  70                  75                  80  
Arg Tyr Asn Pro Ser Leu Lys Ser Arg Ile Ser Ile Thr Arg Asp Thr
                85                  90                  95      
Ser Lys Asn Gln Phe Phe Leu Gln Leu Asn Ser Val Thr Ile Glu Asp
            100                 105                 110         
Thr Ala Thr Tyr Tyr Cys Val Thr Ala Gly Arg Gly Phe Pro Tyr Trp
        115                 120                 125             
Gly Gln Gly Thr Leu Val Thr Val Ser Ala Gly Ser Thr Ser Gly Ser
    130                 135                 140                 
Gly Lys Pro Gly Ser Gly Glu Gly Ser Thr Lys Gly Asp Ile Leu Met
145                 150                 155                 160 
Thr Gln Ser Pro Ser Ser Met Ser Val Ser Leu Gly Asp Thr Val Ser
                165                 170                 175     
Ile Thr Cys His Ser Ser Gln Asp Ile Asn Ser Asn Ile Gly Trp Leu
            180                 185                 190         
Gln Gln Arg Pro Gly Lys Ser Phe Lys Gly Leu Ile Tyr His Gly Thr
        195                 200                 205             
Asn Leu Asp Asp Glu Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly
    210                 215                 220                 
Ala Asp Tyr Ser Leu Thr Ile Ser Ser Leu Glu Ser Glu Asp Phe Ala
225                 230                 235                 240 
Asp Tyr Tyr Cys Val Gln Tyr Ala Gln Phe Pro Trp Thr Phe Gly Gly
                245                 250                 255     
Gly Thr Lys Leu Glu Ile Lys Arg Glu Ser Lys Tyr Gly Pro Pro Cys
            260                 265                 270         
Pro Pro Cys Pro Met Phe Trp Val Leu Val Val Val Gly Gly Val Leu
        275                 280                 285             
Ala Cys Tyr Ser Leu Leu Val Thr Val Ala Phe Ile Ile Phe Trp Val
    290                 295                 300                 
Arg Ser Lys Arg Ser Arg Gly Gly His Ser Asp Tyr Met Asn Met Thr
305                 310                 315                 320 
Pro Arg Arg Pro Gly Pro Thr Arg Lys His Tyr Gln Pro Tyr Ala Pro
                325                 330                 335     
Pro Arg Asp Phe Ala Ala Tyr Arg Ser Arg Val Lys Phe Ser Arg Ser
            340                 345                 350         
Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu
        355                 360                 365             
Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg
    370                 375                 380                 
Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln
385                 390                 395                 400 
Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr
                405                 410                 415     
Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp
            420                 425                 430         
Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala
        435                 440                 445             
Leu His Met Gln Ala Leu Pro Pro Arg Leu Glu Gly Ser Gly Glu Gly
    450                 455                 460                 
Arg Gly Ser Leu Leu Thr Cys Gly Asp Val Glu Glu Asn Pro Gly Pro
465                 470                 475                 480 
Met Leu Leu Leu Val Thr Ser Leu Leu Leu Cys Glu Leu Pro His Pro
                485                 490                 495     
Ala Phe Leu Leu Ile Pro Arg Lys Val Cys Asn Gly Ile Gly Ile Gly
            500                 505                 510         
Glu Phe Lys Asp Ser Leu Ser Ile Asn Ala Thr Asn Ile Lys His Phe
        515                 520                 525             
Lys Asn Cys Thr Ser Ile Ser Gly Asp Leu His Ile Leu Pro Val Ala
    530                 535                 540                 
Phe Arg Gly Asp Ser Phe Thr His Thr Pro Pro Leu Asp Pro Gln Glu
545                 550                 555                 560 
Leu Asp Ile Leu Lys Thr Val Lys Glu Ile Thr Gly Phe Leu Leu Ile
                565                 570                 575     
Gln Ala Trp Pro Glu Asn Arg Thr Asp Leu His Ala Phe Glu Asn Leu
            580                 585                 590         
Glu Ile Ile Arg Gly Arg Thr Lys Gln His Gly Gln Phe Ser Leu Ala
        595                 600                 605             
Val Val Ser Leu Asn Ile Thr Ser Leu Gly Leu Arg Ser Leu Lys Glu
    610                 615                 620                 
Ile Ser Asp Gly Asp Val Ile Ile Ser Gly Asn Lys Asn Leu Cys Tyr
625                 630                 635                 640 
Ala Asn Thr Ile Asn Trp Lys Lys Leu Phe Gly Thr Ser Gly Gln Lys
                645                 650                 655     
Thr Lys Ile Ile Ser Asn Arg Gly Glu Asn Ser Cys Lys Ala Thr Gly
            660                 665                 670         
Gln Val Cys His Ala Leu Cys Ser Pro Glu Gly Cys Trp Gly Pro Glu
        675                 680                 685             
Pro Arg Asp Cys Val Ser Cys Arg Asn Val Ser Arg Gly Arg Glu Cys
    690                 695                 700                 
Val Asp Lys Cys Asn Leu Leu Glu Gly Glu Pro Arg Glu Phe Val Glu
705                 710                 715                 720 
Asn Ser Glu Cys Ile Gln Cys His Pro Glu Cys Leu Pro Gln Ala Met
                725                 730                 735     
Asn Ile Thr Cys Thr Gly Arg Gly Pro Asp Asn Cys Ile Gln Cys Ala
            740                 745                 750         
His Tyr Ile Asp Gly Pro His Cys Val Lys Thr Cys Pro Ala Gly Val
        755                 760                 765             
Met Gly Glu Asn Asn Thr Leu Val Trp Lys Tyr Ala Asp Ala Gly His
    770                 775                 780                 
Val Cys His Leu Cys His Pro Asn Cys Thr Tyr Gly Cys Thr Gly Pro
785                 790                 795                 800 
Gly Leu Glu Gly Cys Pro Thr Asn Gly Pro Lys Ile Pro Ser Ile Ala
                805                 810                 815     
Thr Gly Met Val Gly Ala Leu Leu Leu Leu Leu Val Val Ala Leu Gly
            820                 825                 830         
Ile Gly Leu Phe Met Gly Ser Gly Ala Thr Asn Phe Ser Leu Leu Lys
        835                 840                 845             
Gln Ala Gly Asp Val Glu Glu Asn Pro Gly Pro Met Leu Leu Leu Val
    850                 855                 860                 
Thr Ser Leu Leu Leu Cys Glu Leu Pro His Pro Ala Phe Leu Leu Ile
865                 870                 875                 880 
Pro Ser Val Leu Thr Gln Pro Ser Ser Val Ser Ala Ala Pro Gly Gln
                885                 890                 895     
Lys Val Thr Ile Ser Cys Ser Gly Ser Thr Ser Asn Ile Gly Asn Asn
            900                 905                 910         
Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu Met
        915                 920                 925             
Ile Tyr Asp Val Ser Lys Arg Pro Ser Gly Val Pro Asp Arg Phe Ser
    930                 935                 940                 
Gly Ser Lys Ser Gly Asn Ser Ala Ser Leu Asp Ile Ser Gly Leu Gln
945                 950                 955                 960 
Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu
                965                 970                 975     
Ser Glu Phe Leu Phe Gly Thr Gly Thr Lys Leu Thr Val Leu Gly Gly
            980                 985                 990         
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val
        995                 1000                1005            
Gln Leu Val Glu Ser Gly Gly Asn Leu Val Gln Pro Gly Gly Ser Leu
    1010                1015                1020                
Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Gly Ser Phe Ser Met
1025                1030                1035               1040 
Ser Trp Val Arg Gln Ala Pro Gly Gly Gly Leu Glu Trp Val Ala Gly
                1045                1050                1055    
Leu Ser Ala Arg Ser Ser Leu Thr His Tyr Ala Asp Ser Val Lys Gly
            1060                1065                1070        
Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Val Tyr Leu Gln
        1075                1080                1085            
Met Asn Ser Leu Arg Val Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg
    1090                1095                1100                
Arg Ser Tyr Asp Ser Ser Gly Tyr Trp Gly His Phe Ala Ser Tyr Met
1105                1110                1115               1120 
Asp Val Trp Gly Gln Gly Thr Leu Val Thr Val Ser Glu Ser Lys Tyr
                1125                1130                1135    
Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Asp Gly Gly Pro
            1140                1145                1150        
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
        1155                1160                1165            
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp
    1170                1175                1180                
Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
1185                1190                1195               1200 
Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Gln Ser Thr Tyr Arg Val
                1205                1210                1215    
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
            1220                1225                1230        
Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys
        1235                1240                1245            
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
    1250                1255                1260                
Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr
1265                1270                1275               1280 
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
                1285                1290                1295    
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
            1300                1305                1310        
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys
        1315                1320                1325            
Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu
    1330                1335                1340                
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly
1345                1350                1355               1360 
Lys Met Phe Trp Val Leu Val Val Val Gly Gly Val Leu Ala Cys Tyr
                1365                1370                1375    
Ser Leu Leu Val Thr Val Ala Phe Ile Ile Phe Trp Val Lys Arg Gly
            1380                1385                1390        
Arg Lys Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro Val
        1395                1400                1405            
Gln Thr Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu Glu
    1410                1415                1420                
Glu Glu Gly Gly Cys Glu Leu Arg Val Lys Phe Ser Arg Ser Ala Asp
1425                1430                1435               1440 
Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn
                1445                1450                1455    
Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg
            1460                1465                1470        
Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly
        1475                1480                1485            
Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu
    1490                1495                1500                
Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu
1505                1510                1515               1520 
Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His
                1525                1530                1535    
Met Gln Ala Leu Pro Pro Arg Gly Ser Gly Glu Gly Arg Gly Ser Leu
            1540                1545                1550        
Leu Thr Cys Gly Asp Val Glu Glu Asn Pro Gly Pro Met Val Gly Ser
        1555                1560                1565            
Leu Asn Cys Ile Val Ala Val Ser Gln Asn Met Gly Ile Gly Lys Asn
    1570                1575                1580                
Gly Asp Phe Pro Trp Pro Pro Leu Arg Asn Glu Ser Arg Tyr Phe Gln
1585                1590                1595               1600 
Arg Met Thr Thr Thr Ser Ser Val Glu Gly Lys Gln Asn Leu Val Ile
                1605                1610                1615    
Met Gly Lys Lys Thr Trp Phe Ser Ile Pro Glu Lys Asn Arg Pro Leu
            1620                1625                1630        
Lys Gly Arg Ile Asn Leu Val Leu Ser Arg Glu Leu Lys Glu Pro Pro
        1635                1640                1645            
Gln Gly Ala His Phe Leu Ser Arg Ser Leu Asp Asp Ala Leu Lys Leu
    1650                1655                1660                
Thr Glu Gln Pro Glu Leu Ala Asn Lys Val Asp Met Val Trp Ile Val
1665                1670                1675               1680 
Gly Gly Ser Ser Val Tyr Lys Glu Ala Met Asn His Pro Gly His Leu
                1685                1690                1695    
Lys Leu Phe Val Thr Arg Ile Met Gln Asp Phe Glu Ser Asp Thr Phe
            1700                1705                1710        
Phe Pro Glu Ile Asp Leu Glu Lys Tyr Lys Leu Leu Pro Glu Tyr Pro
        1715                1720                1725            
Gly Val Leu Ser Asp Val Gln Glu Glu Lys Gly Ile Lys Tyr Lys Phe
    1730                1735                1740                
Glu Val Tyr Glu Lys Asn Asp
1745                1750    

