                         SEQUENCE LISTING

<110>  The United States of America, as represented by the 
       Secretary, Department of Health and Human Services; 
       Government of the United States, as Represented by the 
       Secretary of the Army
       Lee, Che-Hung Robert
       Pinto, Valerian B
 
<120>  MULTIVALENT MENINGOCOCCAL CONJUGATES AND METHODS FOR PREPARING 
       CONJUGATES

<130>  4239-89011-02

<150>  US 61/651,382
<151>  2012-05-24

<160>  16    

<170>  PatentIn version 3.5

<210>  1
<211>  525
<212>  DNA
<213>  Neisseria meningitidis

<400>  1
tcagaatttg acgcgcacac cggcggacag ttcgccggaa cggacgtttt tgacagtgtt       60

gactttgccg atgtagttgt agcggtagcc ggcatccaaa tcgacattcg gggtaacggc      120

atagcttacg cccgtcaata cgccgaggcc gatggaggtt tggctgaagc tgtcgctgcc      180

gcccaagtcg acggaggcgc ggttgaggct caagcgcgcg ccgagatacg gtttgacggg      240

cgattgggtg tcgaagtcgt aaatggcgga cgcgccgatg ctgtaaagtt tgaaatcggt      300

ggatggggct ttatagtttt tgtagcgcgt gtaatcgacg gcgaagcgga ggtcgttgat      360

gcggtagcct gcggagatgc gcgggctgaa gcctttggca gaacctaaag agcttgaggc      420

ttttgcgtgt gcggcatcgg cttggacgta aaagccggat gcgccttccg ccagtgcggc      480

ggccgggaga gcgagggcaa tcagtgtggc aagtgctttt ttcat                      525


<210>  2
<211>  174
<212>  PRT
<213>  Neisseria meningitidis

<400>  2

Met Lys Lys Ala Leu Ala Thr Leu Ile Ala Leu Ala Leu Pro Ala Ala 
1               5                   10                  15      


Ala Leu Ala Glu Gly Ala Ser Gly Phe Tyr Val Gln Ala Asp Ala Ala 
            20                  25                  30          


His Ala Lys Ala Ser Ser Ser Leu Gly Ser Ala Lys Gly Phe Ser Pro 
        35                  40                  45              


Arg Ile Ser Ala Gly Tyr Arg Ile Asn Asp Leu Arg Phe Ala Val Asp 
    50                  55                  60                  


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


Tyr Ser Ile Gly Ala Ser Ala Ile Tyr Asp Phe Asp Thr Gln Ser Pro 
                85                  90                  95      


Val Lys Pro Tyr Leu Gly Ala Arg Leu Ser Leu Asn Arg Ala Ser Val 
            100                 105                 110         


Asp Leu Gly Gly Ser Asp Ser Phe Ser Gln Thr Ser Ile Gly Leu Gly 
        115                 120                 125             


Val Leu Thr Gly Val Ser Tyr Ala Val Thr Pro Asn Val Asp Leu Asp 
    130                 135                 140                 


Ala Gly Tyr Arg Tyr Asn Tyr Ile Gly Lys Val Asn Thr Val Lys Asn 
145                 150                 155                 160 


Val Arg Ser Gly Glu Leu Ser Ala Gly Val Arg Val Lys Phe 
                165                 170                 


<210>  3
<211>  765
<212>  DNA
<213>  Neisseria meningitidis

<400>  3
agcagtggag ggggtggtgt cgccgccgac atcggtgcgg ggcttgccga tgcactaacc       60

gcaccgctcg accataaaga caaaggtttg cagtctttga cgctggatca gtccgtcagg      120

aaaaacgaga aactgaagct ggcggcacaa ggtgcggaaa aaacttatgg aaacggtgac      180

agcctcaata cgggcaaatt gaagaacgac aaggtcagcc gtttcgactt tatccgccaa      240

atcgaagtgg acgggcagct cattaccttg gagagtggag agttccaagt atacaaacaa      300

agccattccg ccttaaccgc ctttcagacc gagcaaatac aagattcgga gcattccggg      360

aagatggttg cgaaacgcca gttcagaatc ggcgacatag cgggcgaaca tacatctttt      420

gacaagcttc ccgaaggcgg cagggcgaca tatcgcggga cggcgttcgg ttcagacgat      480

gccggcggaa aactgaccta caccatagat ttcgccgcca agcagggaaa cggcaaaatc      540

gaacatttga aatcgccaga actcaatgtc gacctggccg ccgccgatat caagccggat      600

ggaaaacgcc atgccgtcat cagcggttcc gtcctttaca accaagccga gaaaggcagt      660

tactccctcg gtatctttgg cggaaaagcc caggaagttg ccggcagcgc ggaagtgaaa      720

accgtaaacg gcatacgcca tatcggcctt gccgccaagc aataa                      765


<210>  4
<211>  253
<212>  PRT
<213>  Neisseria meningitidis

<400>  4

Ser Ser Gly Gly Gly Gly Val Ala Ala Asp Ile Gly Ala Gly Leu Ala 
1               5                   10                  15      


Asp Ala Leu Thr Ala Pro Asp His Lys Asp Lys Gly Leu Gln Ser Leu 
            20                  25                  30          


Thr Leu Asp Gln Ser Val Arg Lys Asn Glu Lys Leu Lys Leu Ala Ala 
        35                  40                  45              


Gln Gly Ala Glu Lys Thr Tyr Gly Asn Gly Asp Ser Leu Asn Thr Gly 
    50                  55                  60                  


Lys Leu Lys Asn Asp Lys Val Ser Arg Phe Asp Phe Ile Arg Gln Ile 
65                  70                  75                  80  


Glu Val Asp Gly Gln Leu Ile Thr Leu Glu Ser Gly Glu Phe Gln Val 
                85                  90                  95      


Tyr Lys Gln Ser His Ser Ala Leu Thr Ala Phe Gln Thr Glu Gln Ile 
            100                 105                 110         


Gln Asp Ser Glu His Ser Gly Lys Met Val Ala Lys Arg Gln Phe Arg 
        115                 120                 125             


Ile Gly Asp Ile Ala Gly Glu His Thr Ser Phe Asp Lys Leu Pro Glu 
    130                 135                 140                 


Gly Gly Arg Ala Thr Tyr Arg Gly Thr Ala Phe Gly Ser Asp Asp Ala 
145                 150                 155                 160 


Gly Gly Lys Leu Thr Tyr Thr Ile Asp Phe Ala Ala Lys Gln Gly Asn 
                165                 170                 175     


Gly Lys Ile Glu His Leu Lys Ser Pro Glu Leu Asn Val Asp Leu Ala 
            180                 185                 190         


Ala Ala Asp Ile Lys Pro Asp Gly Lys Arg His Ala Val Ile Ser Gly 
        195                 200                 205             


Ser Val Leu Tyr Asn Gln Ala Glu Lys Gly Ser Tyr Ser Leu Gly Ile 
    210                 215                 220                 


Phe Gly Gly Lys Ala Gln Glu Val Ala Gly Ser Ala Glu Val Lys Thr 
225                 230                 235                 240 


Val Asn Gly Ile Arg His Ile Gly Leu Ala Ala Lys Gln 
                245                 250             


<210>  5
<211>  762
<212>  DNA
<213>  Neisseria meningitidis

<400>  5
agcagtggag gcggcggtgt cgccgccgac atcggcgcgg ggcttgccga tgcactaacc       60

gcaccgctcg accataaaga caaaagtttg cagtctttga cgctggatca gtccgtcagg      120

aaaaacgaga aactgaagct ggcggcacaa ggtgcggaaa aaacttatgg aaacggcgac      180

agcctcaata cgggcaaatt gaagaacgac aaggtcagcc gcttcgactt tatccgtcaa      240

atcgaagtgg acgggcagct cattaccttg gagagcggag agttccaaat atacaaacag      300

gaccactccg ccgtcgttgc cctacagatt gaaaaaatca acaaccccga caaaatcgac      360

agcctgataa accaacgctc cttccttgtc agcggtttgg gcggagaaca taccgccttc      420

aaccaactgc ctgacggcaa agccgagtat cacggcaaag cattcagctc cgacgatgct      480

ggcggaaaac tgacctatac catagatttc gccgccaaac agggacacgg caaaatcgaa      540

cacctgaaaa cacccgagca aaatgtcgag cttgccgccg ccgaactcaa agcagatgaa      600

aaatcacacg ccgtcatttt gggcgacacg cgctacggca gcgaagaaaa aggcacttac      660

cacctcgccc ttttcggcga ccgcgcccaa gaaatcgccg gctcggcaac cgtgaagata      720

ggggaaaagg ttcacgaaat cggcatcgcc ggcaaacagt ag                         762


<210>  6
<211>  252
<212>  PRT
<213>  Neisseria meningitidis

<400>  6

Ser Ser Gly Gly Gly Gly Val Ala Ala Asp Ile Gly Ala Gly Leu Ala 
1               5                   10                  15      


Asp Ala Leu Thr Ala Pro Asp His Lys Asp Lys Ser Leu Gln Ser Leu 
            20                  25                  30          


Thr Leu Asp Gln Ser Val Arg Lys Asn Glu Lys Leu Lys Leu Ala Ala 
        35                  40                  45              


Gln Gly Ala Glu Lys Thr Tyr Gly Asn Gly Asp Ser Leu Asn Thr Gly 
    50                  55                  60                  


Lys Leu Lys Asn Asp Lys Val Ser Arg Phe Asp Phe Ile Arg Gln Ile 
65                  70                  75                  80  


Glu Val Asp Gly Gln Leu Ile Thr Leu Glu Ser Gly Glu Phe Gln Ile 
                85                  90                  95      


Tyr Lys Gln Asp His Ser Ala Val Val Ala Leu Gln Ile Glu Lys Ile 
            100                 105                 110         


Asn Asn Pro Asp Lys Ile Asp Ser Leu Ile Asn Gln Arg Ser Phe Leu 
        115                 120                 125             


Val Ser Gly Leu Gly Gly Glu His Thr Ala Phe Asn Gln Leu Pro Asp 
    130                 135                 140                 


Gly Lys Ala Glu Tyr His Gly Lys Ala Phe Ser Ser Asp Asp Ala Gly 
145                 150                 155                 160 


Gly Lys Leu Thr Tyr Thr Ile Asp Phe Ala Ala Lys Gln Gly His Gly 
                165                 170                 175     


Lys Ile Glu His Leu Lys Thr Pro Glu Gln Asn Val Glu Leu Ala Ala 
            180                 185                 190         


Ala Glu Leu Lys Ala Asp Glu Lys Ser His Ala Val Ile Leu Gly Asp 
        195                 200                 205             


Thr Arg Tyr Gly Ser Glu Glu Lys Gly Thr Tyr His Leu Ala Leu Phe 
    210                 215                 220                 


Gly Asp Arg Ala Gln Glu Ile Ala Gly Ser Ala Thr Val Lys Ile Gly 
225                 230                 235                 240 


Glu Lys Val His Glu Ile Gly Ile Ala Gly Lys Gln 
                245                 250         


<210>  7
<211>  1563
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  nucelic acid encoding fHbp1-fHbp2 fusion protein

<400>  7
agcagtggag ggggtggtgt cgccgccgac atcggtgcgg ggcttgccga tgcactaacc       60

gcaccgctcg accataaaga caaaggtttg cagtctttga cgctggatca gtccgtcagg      120

aaaaacgaga aactgaagct ggcggcacaa ggtgcggaaa aaacttatgg aaacggtgac      180

agcctcaata cgggcaaatt gaagaacgac aaggtcagcc gtttcgactt tatccgccaa      240

atcgaagtgg acgggcagct cattaccttg gagagtggag agttccaagt atacaaacaa      300

agccattccg ccttaaccgc ctttcagacc gagcaaatac aagattcgga gcattccggg      360

aagatggttg cgaaacgcca gttcagaatc ggcgacatag cgggcgaaca tacatctttt      420

gacaagcttc ccgaaggcgg cagggcgaca tatcgcggga cggcgttcgg ttcagacgat      480

gccggcggaa aactgaccta caccatagat ttcgccgcca agcagggaaa cggcaaaatc      540

gaacatttga aatcgccaga actcaatgtc gacctggccg ccgccgatat caagccggat      600

ggaaaacgcc atgccgtcat cagcggttcc gtcctttaca accaagccga gaaaggcagt      660

tactccctcg gtatctttgg cggaaaagcc caggaagttg ccggcagcgc ggaagtgaaa      720

accgtaaacg gcatacgcca tatcggcctt gccaagcaag gtggaggcgg ttcaggcgga      780

ggtggctctg gcggtggcgg ttctagagga ggcggcggtg tcgccgccga catcggcgcg      840

gggcttgccg atgcactaac cgcaccgctc gaccataaag acaaaagttt gcagtctttg      900

acgctggatc agtccgtcag gaaaaacgag aaactgaagc tggcggcaca aggtgcggaa      960

aaaacttatg gaaacggcga cagcctcaat acgggcaaat tgaagaacga caaggtcagc     1020

cgcttcgact ttatccgtca aatcgaagtg gacgggcagc tcattacctt ggagagcgga     1080

gagttccaaa tatacaaaca ggaccactcc gccgtcgttg ccctacagat tgaaaaaatc     1140

aacaaccccg acaaaatcga cagcctgata aaccaacgct ccttccttgt cagcggtttg     1200

ggcggagaac ataccgcctt caaccaactg cctgacggca aagccgagta tcacggcaaa     1260

gcattcagct ccgacgatgc tggcggaaaa ctgacctata ccatagattt cgccgccaaa     1320

cagggacacg gcaaaatcga acacctgaaa acacccgagc aaaatgtcga gcttgccgcc     1380

gccgaactca aagcagatga aaaatcacac gccgtcattt tgggcgacac gcgctacggc     1440

agcgaagaaa aaggcactta ccacctcgcc cttttcggcg accgcgccca agaaatcgcc     1500

ggctcggcaa ccgtgaagat aggggaaaag gttcacgaaa tcggcatcgc cggcaaacag     1560

tag                                                                   1563


<210>  8
<211>  520
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  fHbp1-fHbp2 fusion protein

<400>  8

Ser Ser Gly Gly Gly Gly Val Ala Ala Asp Ile Gly Ala Gly Leu Ala 
1               5                   10                  15      


Asp Ala Leu Thr Ala Pro Asp His Lys Asp Lys Gly Leu Gln Ser Leu 
            20                  25                  30          


Thr Leu Asp Gln Ser Val Arg Lys Asn Glu Lys Leu Lys Leu Ala Ala 
        35                  40                  45              


Gln Gly Ala Glu Lys Thr Tyr Gly Asn Gly Asp Ser Leu Asn Thr Gly 
    50                  55                  60                  


Lys Leu Lys Asn Asp Lys Val Ser Arg Phe Asp Phe Ile Arg Gln Ile 
65                  70                  75                  80  


Glu Val Asp Gly Gln Leu Ile Thr Leu Glu Ser Gly Glu Phe Gln Val 
                85                  90                  95      


Tyr Lys Gln Ser His Ser Ala Leu Thr Ala Phe Gln Thr Glu Gln Ile 
            100                 105                 110         


Gln Asp Ser Glu His Ser Gly Lys Met Val Ala Lys Arg Gln Phe Arg 
        115                 120                 125             


Ile Gly Asp Ile Ala Gly Glu His Thr Ser Phe Asp Lys Leu Pro Glu 
    130                 135                 140                 


Gly Gly Arg Ala Thr Tyr Arg Gly Thr Ala Phe Gly Ser Asp Asp Ala 
145                 150                 155                 160 


Gly Gly Lys Leu Thr Tyr Thr Ile Asp Phe Ala Ala Lys Gln Gly Asn 
                165                 170                 175     


Gly Lys Ile Glu His Leu Lys Ser Pro Glu Leu Asn Val Asp Leu Ala 
            180                 185                 190         


Ala Ala Asp Ile Lys Pro Asp Gly Lys Arg His Ala Val Ile Ser Gly 
        195                 200                 205             


Ser Val Leu Tyr Asn Gln Ala Glu Lys Gly Ser Tyr Ser Leu Gly Ile 
    210                 215                 220                 


Phe Gly Gly Lys Ala Gln Glu Val Ala Gly Ser Ala Glu Val Lys Thr 
225                 230                 235                 240 


Val Asn Gly Ile Arg His Ile Gly Leu Ala Lys Gln Gly Gly Gly Gly 
                245                 250                 255     


Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Arg Ser Ser Gly Gly 
            260                 265                 270         


Gly Gly Val Ala Ala Asp Ile Gly Ala Gly Leu Ala Asp Ala Leu Thr 
        275                 280                 285             


Ala Pro Asp His Lys Asp Lys Ser Leu Gln Ser Leu Thr Leu Asp Gln 
    290                 295                 300                 


Ser Val Arg Lys Asn Glu Lys Leu Lys Leu Ala Ala Gln Gly Ala Glu 
305                 310                 315                 320 


Lys Thr Tyr Gly Asn Gly Asp Ser Leu Asn Thr Gly Lys Leu Lys Asn 
                325                 330                 335     


Asp Lys Val Ser Arg Phe Asp Phe Ile Arg Gln Ile Glu Val Asp Gly 
            340                 345                 350         


Gln Leu Ile Thr Leu Glu Ser Gly Glu Phe Gln Ile Tyr Lys Gln Asp 
        355                 360                 365             


His Ser Ala Val Val Ala Leu Gln Ile Glu Lys Ile Asn Asn Pro Asp 
    370                 375                 380                 


Lys Ile Asp Ser Leu Ile Asn Gln Arg Ser Phe Leu Val Ser Gly Leu 
385                 390                 395                 400 


Gly Gly Glu His Thr Ala Phe Asn Gln Leu Pro Asp Gly Lys Ala Glu 
                405                 410                 415     


Tyr His Gly Lys Ala Phe Ser Ser Asp Asp Ala Gly Gly Lys Leu Thr 
            420                 425                 430         


Tyr Thr Ile Asp Phe Ala Ala Lys Gln Gly His Gly Lys Ile Glu His 
        435                 440                 445             


Leu Lys Thr Pro Glu Gln Asn Val Glu Leu Ala Ala Ala Glu Leu Lys 
    450                 455                 460                 


Ala Asp Glu Lys Ser His Ala Val Ile Leu Gly Asp Thr Arg Tyr Gly 
465                 470                 475                 480 


Ser Glu Glu Lys Gly Thr Tyr His Leu Ala Leu Phe Gly Asp Arg Ala 
                485                 490                 495     


Gln Glu Ile Ala Gly Ser Ala Thr Val Lys Ile Gly Glu Lys Val His 
            500                 505                 510         


Glu Ile Gly Ile Ala Gly Lys Gln 
        515                 520 


<210>  9
<211>  39
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic oligonucleotide primer

<400>  9
aagcttcctc gagtgagcag tggagggggt ggtgtcgcc                              39


<210>  10
<211>  38
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic oligonucleotide primer

<400>  10
ggcggggaat tcacttattg cttggcggca aggccgat                               38


<210>  11
<211>  39
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic oligonucleotide primer

<400>  11
aagcttcctc gagtgagcag tggaggcggc ggtgtcgcc                              39


<210>  12
<211>  38
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic oligonucleotide primer

<400>  12
ggcggggaat tcacttacta ctgtttgccg gcgatgcc                               38


<210>  13
<211>  63
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic oligonucleotide primer

<400>  13
cgatccgcca ccgccagagc cacctccgcc tgaaccgcct ccaccttgct tggcaaggcc       60

gat                                                                     63


<210>  14
<211>  36
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic oligonucleotide primer

<400>  14
gccgcctcct ctagaaccgc caccgccaga gccacc                                 36


<210>  15
<211>  63
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic oligonucleotide primer

<400>  15
ggtggaggcg gttcaggcgg aggtggctct ggcggtggcg gatcgggagg cggcggtgtc       60

gcc                                                                     63


<210>  16
<211>  33
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic oligonucleotide primer

<400>  16
ggtggcggtt ctagaggagg cggcggtgtc gcc                                    33


