                         SEQUENCE LISTING

<110>  INTELECTYS
 
<120>  ANTIBODIES AND FRAGMENTS THEREOF RAISED AGAINST THE ALPHA-3 
       DOMAIN OF HLA-G PROTEIN, METHODS AND MEANS FOR THEIR PREPARATION,
        AND USES THEREOF

<130>  B10037A - AD/AFL/KN

<140>  PCT/EP XXXXXX
<141>  2013-11-12

<150>  EP12306398.4
<151>  2012-11-12

<160>  12    

<170>  PatentIn version 3.5

<210>  1
<211>  92
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  HLA-Galpha3 protein sequence

<400>  1

Asp Pro Pro Lys Thr His Val Thr His His Pro Val Phe Asp Tyr Glu 
1               5                   10                  15      


Ala Thr Leu Arg Cys Trp Ala Leu Gly Phe Tyr Pro Ala Glu Ile Ile 
            20                  25                  30          


Leu Thr Trp Gln Arg Asp Gly Glu Asp Gln Thr Gln Asp Val Glu Leu 
        35                  40                  45              


Val Glu Thr Arg Pro Ala Gly Asp Gly Thr Phe Gln Lys Trp Ala Ala 
    50                  55                  60                  


Val Val Val Pro Ser Gly Glu Glu Gln Arg Tyr Thr Cys His Val Gln 
65                  70                  75                  80  


His Glu Gly Leu Pro Glu Pro Leu Met Leu Arg Trp 
                85                  90          


<210>  2
<211>  108
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  HLA-G ALPHA-3 domain used for protein immunization


<220>
<221>  SITE
<222>  (1)..(12)
<223>  linker sequence as found under SEQ ID N0 3

<400>  2

Gly Cys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Arg Ala Asp Pro 
1               5                   10                  15      


Pro Lys Thr His Val Thr His His Pro Val Phe Asp Tyr Glu Ala Thr 
            20                  25                  30          


Leu Arg Cys Trp Ala Leu Gly Phe Tyr Pro Ala Glu Ile Ile Leu Thr 
        35                  40                  45              


Trp Gln Arg Asp Gly Glu Asp Gln Thr Gln Asp Val Glu Leu Val Glu 
    50                  55                  60                  


Thr Arg Pro Ala Gly Asp Gly Thr Phe Gln Lys Trp Ala Ala Val Val 
65                  70                  75                  80  


Val Pro Ser Gly Glu Glu Gln Arg Tyr Thr Cys His Val Gln His Glu 
                85                  90                  95      


Gly Leu Pro Glu Pro Leu Met Leu Arg Trp Lys Gln 
            100                 105             


<210>  3
<211>  12
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Linker sequence

<400>  3

Gly Cys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 
1               5                   10          


<210>  4
<211>  337
<212>  PRT
<213>  Homo sapiens


<220>
<221>  MISC_FEATURE
<222>  (1)..(337)
<223>  full HLA-G human protein as found under NM_002127.5

<400>  4

Met Val Val Met Ala Pro Arg Thr Leu Phe Leu Leu Leu Ser Gly Ala 
1               5                   10                  15      


Leu Thr Leu Thr Glu Thr Trp Ala Gly Ser His Ser Met Arg Tyr Phe 
            20                  25                  30          


Ser Ala Ala Val Ser Arg Pro Gly Arg Gly Glu Pro Arg Phe Ile Ala 
        35                  40                  45              


Met Gly Tyr Val Asp Asp Thr Gln Phe Val Arg Phe Asp Ser Asp Ser 
    50                  55                  60                  


Ala Cys Pro Arg Met Glu Pro Arg Ala Pro Trp Val Glu Gln Glu Gly 
65                  70                  75                  80  


Pro Glu Tyr Trp Glu Glu Glu Thr Arg Asn Thr Lys Ala His Ala Gln 
                85                  90                  95      


Thr Asp Arg Met Asn Leu Thr Leu Arg Gly Tyr Tyr Asn Gln Ser Glu 
            100                 105                 110         


Ala Ser Ser His Thr Leu Gln Trp Met Ile Gly Cys Asp Leu Gly Ser 
        115                 120                 125             


Asp Gly Arg Leu Leu Arg Gly Tyr Glu Gln Tyr Ala Tyr Asp Gly Lys 
    130                 135                 140                 


Asp Tyr Leu Ala Leu Asn Glu Asp Leu Arg Ser Trp Thr Ala Ala Asp 
145                 150                 155                 160 


Thr Ala Ala Gln Ile Ser Lys Arg Lys Cys Glu Ala Ala Asn Val Ala 
                165                 170                 175     


Glu Gln Arg Arg Ala Tyr Leu Glu Gly Thr Cys Val Glu Trp Leu His 
            180                 185                 190         


Arg Tyr Leu Glu Asn Gly Lys Glu Met Leu Gln Arg Ala Asp Pro Pro 
        195                 200                 205             


Lys Thr His Val Thr His His Pro Val Phe Asp Tyr Glu Ala Thr Leu 
    210                 215                 220                 


Arg Cys Trp Ala Leu Gly Phe Tyr Pro Ala Glu Ile Ile Leu Thr Trp 
225                 230                 235                 240 


Gln Arg Asp Gly Glu Asp Gln Thr Gln Asp Val Glu Leu Val Glu Thr 
                245                 250                 255     


Arg Pro Ala Gly Asp Gly Thr Phe Gln Lys Trp Ala Ala Val Val Val 
            260                 265                 270         


Pro Ser Gly Glu Glu Gln Arg Tyr Thr Cys His Val Gln His Glu Gly 
        275                 280                 285             


Leu Pro Glu Pro Leu Met Leu Arg Trp Lys Gln Ser Ser Leu Pro Thr 
    290                 295                 300                 


Ile Pro Ile Met Gly Ile Val Ala Gly Leu Val Val Leu Ala Ala Val 
305                 310                 315                 320 


Val Thr Gly Ala Ala Val Ala Ala Val Leu Trp Arg Lys Lys Ser Ser 
                325                 330                 335     


Asp 
    


<210>  5
<211>  1578
<212>  DNA
<213>  Homo sapiens


<220>
<221>  source
<222>  (1)..(1578)
<223>  full human HLA-G nucleic acid sequence as found under NM_002127.5

<400>  5
agtgtggtac tttgtcttga ggagatgtcc tggactcaca cggaaactta gggctacgga       60

atgaagttct cactcccatt aggtgacagg tttttagaga agccaatcag cgtcgccgcg      120

gtcctggttc taaagtcctc gctcacccac ccggactcat tctccccaga cgccaaggat      180

ggtggtcatg gcgccccgaa ccctcttcct gctgctctcg ggggccctga ccctgaccga      240

gacctgggcg ggctcccact ccatgaggta tttcagcgcc gccgtgtccc ggcccggccg      300

cggggagccc cgcttcatcg ccatgggcta cgtggacgac acgcagttcg tgcggttcga      360

cagcgactcg gcgtgtccga ggatggagcc gcgggcgccg tgggtggagc aggaggggcc      420

ggagtattgg gaagaggaga cacggaacac caaggcccac gcacagactg acagaatgaa      480

cctgcagacc ctgcgcggct actacaacca gagcgaggcc agttctcaca ccctccagtg      540

gatgattggc tgcgacctgg ggtccgacgg acgcctcctc cgcgggtatg aacagtatgc      600

ctacgatggc aaggattacc tcgccctgaa cgaggacctg cgctcctgga ccgcagcgga      660

cactgcggct cagatctcca agcgcaagtg tgaggcggcc aatgtggctg aacaaaggag      720

agcctacctg gagggcacgt gcgtggagtg gctccacaga tacctggaga acgggaagga      780

gatgctgcag cgcgcggacc cccccaagac acacgtgacc caccaccctg tctttgacta      840

tgaggccacc ctgaggtgct gggccctggg cttctaccct gcggagatca tactgacctg      900

gcagcgggat ggggaggacc agacccagga cgtggagctc gtggagacca ggcctgcagg      960

ggatggaacc ttccagaagt gggcagctgt ggtggtgcct tctggagagg agcagagata     1020

cacgtgccat gtgcagcatg aggggctgcc ggagcccctc atgctgagat ggaagcagtc     1080

ttccctgccc accatcccca tcatgggtat cgttgctggc ctggttgtcc ttgcagctgt     1140

agtcactgga gctgcggtcg ctgctgtgct gtggagaaag aagagctcag attgaaaagg     1200

agggagctac tctcaggctg caatgtgaaa cagctgccct gtgtgggact gagtggcaag     1260

tccctttgtg acttcaagaa ccctgactcc tctttgtgca gagaccagcc cacccctgtg     1320

cccaccatga ccctcttcct catgctgaac tgcattcctt ccccaatcac ctttcctgtt     1380

ccagaaaagg ggctgggatg tctccgtctc tgtctcaaat ttgtggtcca ctgagctata     1440

acttacttct gtattaaaat tagaatctga gtataaattt actttttcaa attatttcca     1500

agagagattg atgggttaat taaaggagaa gattcctgaa atttgagaga caaaataaat     1560

ggaagacatg agaacttt                                                   1578


<210>  6
<211>  546
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence used for DNA immunization


<220>
<221>  misc_feature
<222>  (1)..(6)
<223>  Restriction site

<220>
<221>  misc_feature
<222>  (7)..(15)
<223>  5' kozak sequence (SEQ ID N7)

<220>
<221>  sig_peptide
<222>  (13)..(84)
<223>  Signal peptide as found in Exon 1 of HLA-G

<220>
<221>  misc_feature
<222>  (85)..(87)
<223>  Nucleotides encoding Added amino acids

<220>
<221>  misc_feature
<222>  (88)..(93)
<223>  Nucleotides found at the end of the alpha 2 portion of HLA-G

<220>
<221>  misc_feature
<222>  (94)..(369)
<223>  Alpha 3 domain (SEQ ID NO 10)

<220>
<221>  misc_feature
<222>  (370)..(486)
<223>  Transmembrane-anchor protein encoding portion

<220>
<221>  misc_feature
<222>  (487)..(489)
<223>  nucleotides encoding added amino acids

<220>
<221>  misc_feature
<222>  (490)..(531)
<223>  V5-tag sequence

<220>
<221>  misc_feature
<222>  (532)..(540)
<223>  3' stop codons (SEQ ID NO 8)

<220>
<221>  misc_feature
<222>  (541)..(546)
<223>  Restriction site

<400>  6
aagcttgccg ccatggtcgt tatggcaccc aggaccttgt tcctcctgct ctctggagca       60

ctgaccctta ctgagacatg ggccagtaga gccgaccccc ccaaaaccca tgtgacccac      120

cacccagtct ttgactatga agctacactg agatgttggg ccctgggctt ctaccccgca      180

gagatcatcc tgacctggca gcgcgacgga gaagatcaga cacaggacgt cgagctcgtg      240

gaaacccggc ctgctggtga tggcacattt cagaagtggg ccgccgtggt ggttccatcc      300

ggtgaggaac agcgctacac ttgccatgtg cagcacgagg gcttgcctga gcctcttatg      360

cttcggtgga agcagtcatc cctgccaact attcccatca tgggcattgt ggccggactg      420

gtggttctgg cagctgtggt gactggcgct gccgtcgccg ctgtcctctg gaggaaaaag      480

agcagcgatg gcaagccaat tcctaatcca ttgctgggcc tggactcaac ttgatgataa      540

ctcgag                                                                 546


<210>  7
<211>  9
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  5' kozak sequence

<400>  7
gccgccatg                                                                9


<210>  8
<211>  9
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  3' stop codons

<400>  8
tgatgataa                                                                9


<210>  9
<211>  276
<212>  DNA
<213>  Homo sapiens


<220>
<221>  misc_feature
<222>  (1)..(276)
<223>  nucleic acid sequence encoding a HLA-G ALPHA-3 domain polypeptide
       sequence as found in the sequence disclosed under accession 
       number NM_002127.5, between positions 797 and 1072

<400>  9
gaccccccca agacacacgt gacccaccac cctgtctttg actatgaggc caccctgagg       60

tgctgggccc tgggcttcta ccctgcggag atcatactga cctggcagcg ggatggggag      120

gaccagaccc aggacgtgga gctcgtggag accaggcctg caggggatgg aaccttccag      180

aagtgggcag ctgtggtggt gccttctgga gaggagcaga gatacacgtg ccatgtgcag      240

catgaggggc tgccggagcc cctcatgctg agatgg                                276


<210>  10
<211>  276
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  optimized nucleic acid sequence encoding for an HLA-G ALPHA-3 
       domain polypeptide sequence

<400>  10
gaccccccca aaacccatgt gacccaccac ccagtctttg actatgaagc tacactgaga       60

tgttgggccc tgggcttcta ccccgcagag atcatcctga cctggcagcg cgacggagaa      120

gatcagacac aggacgtcga gctcgtggaa acccggcctg ctggtgatgg cacatttcag      180

aagtgggccg ccgtggtggt tccatccggt gaggaacagc gctacacttg ccatgtgcag      240

cacgagggct tgcctgagcc tcttatgctt cggtgg                                276


<210>  11
<211>  173
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Protein sequence encoded by the nucleic acid sequence SEQ ID NO 6


<220>
<221>  SIGNAL
<222>  (1)..(24)
<223>  Signal peptide as found in Exon 1 of HLA-G

<220>
<221>  SITE
<222>  (25)..(25)
<223>  Added amino-acid

<220>
<221>  SITE
<222>  (26)..(27)
<223>  Nucleotides found at the end of the Alpha2 portion of HLA-G

<220>
<221>  DOMAIN
<222>  (28)..(119)
<223>  Alpha3 domain

<220>
<221>  TRANSMEM
<222>  (120)..(158)
<223>  Transmembrane-anchor protein encoding portion

<220>
<221>  SITE
<222>  (159)..(159)
<223>  Added amino-acid

<220>
<221>  SITE
<222>  (160)..(173)
<223>  V5-tag sequence

<400>  11

Met Val Val Met Ala Pro Arg Thr Leu Phe Leu Leu Leu Ser Gly Ala 
1               5                   10                  15      


Leu Thr Leu Thr Glu Thr Trp Ala Ser Arg Ala Asp Pro Pro Lys Thr 
            20                  25                  30          


His Val Thr His His Pro Val Phe Asp Tyr Glu Ala Thr Leu Arg Cys 
        35                  40                  45              


Trp Ala Leu Gly Phe Tyr Pro Ala Glu Ile Ile Leu Thr Trp Gln Arg 
    50                  55                  60                  


Asp Gly Glu Asp Gln Thr Gln Asp Val Glu Leu Val Glu Thr Arg Pro 
65                  70                  75                  80  


Ala Gly Asp Gly Thr Phe Gln Lys Trp Ala Ala Val Val Val Pro Ser 
                85                  90                  95      


Gly Glu Glu Gln Arg Tyr Thr Cys His Val Gln His Glu Gly Leu Pro 
            100                 105                 110         


Glu Pro Leu Met Leu Arg Trp Lys Gln Ser Ser Leu Pro Thr Ile Pro 
        115                 120                 125             


Ile Met Gly Ile Val Ala Gly Leu Val Val Leu Ala Ala Val Val Thr 
    130                 135                 140                 


Gly Ala Ala Val Ala Ala Val Leu Trp Arg Lys Lys Ser Ser Asp Gly 
145                 150                 155                 160 


Lys Pro Ile Pro Asn Pro Leu Leu Gly Leu Asp Ser Thr 
                165                 170             


<210>  12
<211>  108
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Sequence detailed in figure 5

<400>  12

Gly Cys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Arg Ala Asp Pro 
1               5                   10                  15      


Pro Lys Thr His Val Thr His His Pro Val Phe Asp Tyr Glu Ala Thr 
            20                  25                  30          


Leu Arg Cys Trp Ala Leu Gly Phe Tyr Pro Ala Glu Ile Ile Leu Thr 
        35                  40                  45              


Trp Gln Arg Asp Gly Glu Asp Gln Thr Gln Asp Val Glu Leu Val Glu 
    50                  55                  60                  


Thr Arg Pro Ala Gly Asp Gly Thr Phe Gln Lys Trp Ala Ala Val Val 
65                  70                  75                  80  


Val Pro Ser Gly Glu Glu Gln Arg Tyr Thr Cys His Val Gln His Glu 
                85                  90                  95      


Gly Leu Pro Glu Pro Leu Met Leu Arg Trp Lys Gln 
            100                 105             


