                         SEQUENCE LISTING

<110>  The Johns Hopkins University
 
<120>  IL-6Ralpha/IL-8R BISPECIFIC BINDING AGENTS FOR INHIBITING CANCER 
       CELL MIGRATION

<130>  44807-0323WO1

<150>  62/855,625
<151>  2019-05-31

<160>  49    

<170>  PatentIn version 3.5

<210>  1
<211>  4
<212>  PRT
<213>  Artificial

<220>
<223>  Exemplary Linker

<400>  1

Gly Gly Gly Ser 
1               


<210>  2
<211>  5
<212>  PRT
<213>  Artificial

<220>
<223>  Exemplary Linker

<400>  2

Gly Gly Gly Gly Ser 
1               5   


<210>  3
<211>  4
<212>  PRT
<213>  Artificial

<220>
<223>  Exemplary Linker

<400>  3

Gly Gly Ser Gly 
1               


<210>  4
<211>  14
<212>  PRT
<213>  Artificial

<220>
<223>  Exemplary Linker

<400>  4

Gly Ser Ala Ala Ala Gly Gly Ser Gly Gly Ser Gly Gly Ser 
1               5                   10                  


<210>  5
<211>  8
<212>  PRT
<213>  Artificial

<220>
<223>  Exemplary Linker

<400>  5

Gly Gly Gly Ser Gly Gly Gly Ser 
1               5               


<210>  6
<211>  330
<212>  PRT
<213>  Homo sapiens

<400>  6

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


Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 
            20                  25                  30          


Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 
        35                  40                  45              


Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser 
    50                  55                  60                  


Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr 
65                  70                  75                  80  


Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys 
                85                  90                  95      


Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys 
            100                 105                 110         


Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro 
        115                 120                 125             


Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 
    130                 135                 140                 


Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 
145                 150                 155                 160 


Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 
                165                 170                 175     


Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 
            180                 185                 190         


His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 
        195                 200                 205             


Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 
    210                 215                 220                 


Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu 
225                 230                 235                 240 


Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 
                245                 250                 255     


Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 
            260                 265                 270         


Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 
        275                 280                 285             


Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 
    290                 295                 300                 


Val Phe Ser Cys Ser Val Met His Glu Gly Leu His Asn His Tyr Thr 
305                 310                 315                 320 


Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 
                325                 330 


<210>  7
<211>  330
<212>  PRT
<213>  Homo sapiens

<400>  7

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


Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 
            20                  25                  30          


Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 
        35                  40                  45              


Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser 
    50                  55                  60                  


Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr 
65                  70                  75                  80  


Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys 
                85                  90                  95      


Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys 
            100                 105                 110         


Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro 
        115                 120                 125             


Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 
    130                 135                 140                 


Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 
145                 150                 155                 160 


Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 
                165                 170                 175     


Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 
            180                 185                 190         


His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 
        195                 200                 205             


Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 
    210                 215                 220                 


Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu 
225                 230                 235                 240 


Met Thr Lys Asn Gln Val Ser Leu Ser Cys Ala Val Lys Gly Phe Tyr 
                245                 250                 255     


Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 
            260                 265                 270         


Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 
        275                 280                 285             


Leu Val Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 
    290                 295                 300                 


Val Phe Ser Cys Ser Val Met His Glu Gly Leu His Asn His Tyr Thr 
305                 310                 315                 320 


Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 
                325                 330 


<210>  8
<211>  330
<212>  PRT
<213>  Homo sapiens

<400>  8

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


Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 
            20                  25                  30          


Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 
        35                  40                  45              


Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser 
    50                  55                  60                  


Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr 
65                  70                  75                  80  


Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys 
                85                  90                  95      


Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys 
            100                 105                 110         


Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro 
        115                 120                 125             


Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 
    130                 135                 140                 


Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 
145                 150                 155                 160 


Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 
                165                 170                 175     


Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 
            180                 185                 190         


His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 
        195                 200                 205             


Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 
    210                 215                 220                 


Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu 
225                 230                 235                 240 


Met Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val Lys Gly Phe Tyr 
                245                 250                 255     


Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 
            260                 265                 270         


Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 
        275                 280                 285             


Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 
    290                 295                 300                 


Val Phe Ser Cys Ser Val Met His Glu Gly Leu His Asn His Tyr Thr 
305                 310                 315                 320 


Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 
                325                 330 


<210>  9
<211>  726
<212>  PRT
<213>  Artificial

<220>
<223>  BS1 Antibody Chain 1 with Tociliuzmab Light and Heavy Chains and 
       Human IgG1 CH1, CH2, and CH3 Domains with "Holes" Substitutions

<400>  9

Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp Leu Pro 
1               5                   10                  15      


Gly Ala Arg Cys Ala Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser 
            20                  25                  30          


Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala 
        35                  40                  45              


Ser Gln Asp Ile Ser Ser Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly 
    50                  55                  60                  


Lys Ala Pro Lys Leu Leu Ile Tyr Tyr Thr Ser Arg Leu His Ser Gly 
65                  70                  75                  80  


Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Phe 
                85                  90                  95      


Thr Ile Ser Ser Leu Gln Pro Glu Asp Ile Ala Thr Tyr Tyr Cys Gln 
            100                 105                 110         


Gln Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gln Gly Thr Lys Val Glu 
        115                 120                 125             


Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser 
    130                 135                 140                 


Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn 
145                 150                 155                 160 


Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala 
                165                 170                 175     


Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys 
            180                 185                 190         


Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp 
        195                 200                 205             


Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu 
    210                 215                 220                 


Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys Gly Gly Ser 
225                 230                 235                 240 


Gly Gly Ser Ser Gly Ser Gly Ser Gly Ser Thr Gly Thr Ser Ser Ser 
                245                 250                 255     


Gly Thr Gly Thr Ser Ala Gly Thr Thr Gly Thr Ser Ala Ser Thr Ser 
            260                 265                 270         


Gly Ser Gly Ser Gly Glu Val Gln Leu Gln Glu Ser Gly Pro Gly Leu 
        275                 280                 285             


Val Arg Pro Ser Gln Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Tyr 
    290                 295                 300                 


Ser Ile Thr Ser Asp His Ala Trp Ser Trp Val Arg Gln Pro Pro Gly 
305                 310                 315                 320 


Arg Gly Leu Glu Trp Ile Gly Tyr Ile Ser Tyr Ser Gly Ile Thr Thr 
                325                 330                 335     


Tyr Asn Pro Ser Leu Lys Ser Arg Val Thr Met Leu Arg Asp Thr Ser 
            340                 345                 350         


Lys Asn Gln Phe Ser Leu Arg Leu Ser Ser Val Thr Ala Ala Asp Thr 
        355                 360                 365             


Ala Val Tyr Tyr Cys Ala Arg Ser Leu Ala Arg Thr Thr Ala Met Asp 
    370                 375                 380                 


Tyr Trp Gly Gln Gly Ser Leu Val Thr Val Ser Ser Ala Ser Thr Lys 
385                 390                 395                 400 


Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly 
                405                 410                 415     


Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro 
            420                 425                 430         


Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr 
        435                 440                 445             


Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val 
    450                 455                 460                 


Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn 
465                 470                 475                 480 


Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro 
                485                 490                 495     


Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu 
            500                 505                 510         


Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 
        515                 520                 525             


Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 
    530                 535                 540                 


Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 
545                 550                 555                 560 


Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 
                565                 570                 575     


Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 
            580                 585                 590         


Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 
        595                 600                 605             


Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 
    610                 615                 620                 


Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn 
625                 630                 635                 640 


Gln Val Ser Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile 
                645                 650                 655     


Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 
            660                 665                 670         


Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys 
        675                 680                 685             


Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 
    690                 695                 700                 


Ser Val Met His Glu Gly Leu His Asn His Tyr Thr Gln Lys Ser Leu 
705                 710                 715                 720 


Ser Leu Ser Pro Gly Lys 
                725     


<210>  10
<211>  2178
<212>  DNA
<213>  Artificial

<220>
<223>  Nucleic Acid Sequence Encoding BS1 Antibody Chain 1 with 
       Tocilizumab Light and Heavy Chains and Human IgG1 CH1, CH2, and 
       CH3 Domains with "Holes" Substitutions

<400>  10
atgagggtcc ccgctcagct cctggggctc ctgctgctct ggctcccagg tgcacgatgt       60

gccggatccg acatccagat gacccagagc cccagcagcc tgagcgccag cgtgggcgac      120

cgcgtgacca tcacctgccg cgccagccag gacatcagca gctacctgaa ctggtaccag      180

cagaagcccg gcaaggcccc caagctgctg atctactaca ccagccgcct gcacagcggc      240

gtgcccagcc gcttcagcgg cagcggcagc ggcaccgact tcaccttcac catcagcagc      300

ctgcagcccg aggacatcgc cacctactac tgccagcagg gcaacaccct gccctacacc      360

ttcggccagg gcaccaaggt ggagatcaag cgtacggtgg ctgcaccatc tgtcttcatc      420

ttcccgccat ctgatgagca gttgaaatct ggaactgcct ctgttgtgtg cctgctgaat      480

aacttctatc ccagagaggc caaagtacag tggaaggtgg ataacgccct ccaatcgggt      540

aactcccagg agagtgtcac agagcaggac agcaaggaca gcacctacag cctcagcagc      600

accctgacgc tgagcaaagc agactacgag aaacacaaag tctacgcctg cgaagtcacc      660

catcagggcc tgagctcgcc cgtcacaaag agcttcaaca ggggtgagtg cggtggttct      720

gggggatcta gcggatcggg gtctgggtcg actggtacct cgtcaagcgg gacgggtact      780

agtgctggta ctacgggcac ttcagctagc acctctggct cggggtccgg cgaggtgcag      840

ctgcaggaga gcggccccgg cctggtgcgc cccagccaga ccctgagcct gacctgcacc      900

gtgagcggct acagcatcac cagcgaccac gcctggagct gggtgcgcca gccacctggt      960

cgcggactgg agtggatcgg ctacatcagc tacagcggca tcaccaccta caaccccagc     1020

ctgaagagcc gcgtgaccat gctgcgcgac accagcaaga accagttcag cctgcgcctg     1080

agcagcgtga ccgccgccga caccgccgtg tactactgcg cccgcagcct ggcccgcacc     1140

accgccatgg actactgggg ccagggcagc ctggtgaccg tgagcagcgc tagcaccaag     1200

ggcccatcgg tcttccccct ggcaccctcc tccaagagca cctctggggg cacagcggcc     1260

ctgggctgcc tggtcaagga ctacttcccc gaaccggtga cggtgtcgtg gaactcaggc     1320

gccctgacca gcggcgtgca caccttcccg gctgtcctac agtcctcagg actctactcc     1380

ctcagcagcg tggtgaccgt gccctccagc agcttgggca cccagaccta catctgcaac     1440

gtgaatcaca agcccagcaa caccaaggtg gacaagaaag ttgagcccaa atcttgtgac     1500

aaaactcaca catgcccacc gtgcccagca cctgaactcc tggggggacc gtcagtcttc     1560

ctcttccccc caaaacccaa ggacaccctc atgatctccc ggacccctga ggtcacatgc     1620

gtggtggtgg acgtgagcca cgaagaccct gaggtcaagt tcaactggta cgtggacggc     1680

gtggaggtgc ataatgccaa gacaaagccg cgggaggagc agtacaacag cacgtaccgt     1740

gtggtcagcg tcctcaccgt cctgcaccag gactggctga atggcaagga gtacaagtgc     1800

aaggtctcca acaaagccct cccagccccc atcgagaaaa ccatctccaa agccaaaggg     1860

cagccccgag aaccacaggt gtacaccctg cccccatccc gggaggagat gaccaagaac     1920

caggtcagcc tgtcctgcgc tgtcaaaggc ttctatccca gcgacatcgc cgtggagtgg     1980

gagagcaatg ggcagccgga gaacaactac aagaccacgc ctcccgtgct ggactccgac     2040

ggctccttct tcctcgtgag caagctcacc gtggacaaga gcaggtggca gcaggggaac     2100

gtcttctcat gctccgtgat gcatgagggt ctgcacaacc actacacgca gaagagcctc     2160

tccctgtctc cgggtaaa                                                   2178


<210>  11
<211>  723
<212>  PRT
<213>  Artificial

<220>
<223>  BS1 Antibody Chain 2 with 10H2 Light and Heavy Chains and Human 
       IgG1 CH1, CH2, and CH3 Domains with "Knobs" Substitutions

<400>  11

Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp Leu Pro 
1               5                   10                  15      


Gly Ala Arg Cys Ala Gly Ser Gln Ala Val Val Thr Gln Glu Ser Ala 
            20                  25                  30          


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


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


Pro Asp His Leu Phe Thr Gly Leu Ile Gly Gly Thr Asn Asn Arg Pro 
65                  70                  75                  80  


Pro Gly Val Pro Ala Arg Phe Ser Gly Ser Leu Ile Gly Asp Lys Ala 
                85                  90                  95      


Ala Leu Thr Ile Thr Gly Ala Gln Ile Glu Asp Glu Ala Ile Tyr Phe 
            100                 105                 110         


Cys Ala Leu Trp Tyr Ser Asn His Leu Val Phe Gly Gly Gly Thr Lys 
        115                 120                 125             


Leu Thr Val Leu Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro 
    130                 135                 140                 


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


Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp 
                165                 170                 175     


Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp 
            180                 185                 190         


Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys 
        195                 200                 205             


Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln 
    210                 215                 220                 


Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys Gly 
225                 230                 235                 240 


Gly Ser Gly Gly Ser Ser Gly Ser Gly Ser Gly Ser Thr Gly Thr Ser 
                245                 250                 255     


Ser Ser Gly Thr Gly Thr Ser Ala Gly Thr Thr Gly Thr Ser Ala Ser 
            260                 265                 270         


Thr Ser Gly Ser Gly Ser Gly Glu Val Gln Leu Val Glu Thr Gly Gly 
        275                 280                 285             


Arg Leu Val Gln Pro Lys Gly Ser Leu Lys Leu Ser Cys Ala Val Ser 
    290                 295                 300                 


Gly Ile Thr Phe Lys Thr Asn Ala Met Asn Trp Val Arg Gln Ala Pro 
305                 310                 315                 320 


Gly Lys Gly Leu Glu Trp Val Ala Arg Ile Arg Thr Lys Ser Tyr Asn 
                325                 330                 335     


Tyr Ala Thr Tyr Tyr Ala Asp Ser Val Lys Asp Arg Phe Thr Ile Ser 
            340                 345                 350         


Arg Asp Asp Ser Gln Ser Ile Leu Tyr Leu Gln Met Asn Asn Leu Lys 
        355                 360                 365             


Thr Glu Asp Thr Ala Met Tyr His Cys Val Arg Glu Gly Arg Trp Gly 
    370                 375                 380                 


Gln Gly Thr Leu Val Thr Val Ser Ala Ala Ser Thr Lys Gly Pro Ser 
385                 390                 395                 400 


Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala 
                405                 410                 415     


Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val 
            420                 425                 430         


Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala 
        435                 440                 445             


Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val 
    450                 455                 460                 


Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His 
465                 470                 475                 480 


Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys 
                485                 490                 495     


Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly 
            500                 505                 510         


Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met 
        515                 520                 525             


Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His 
    530                 535                 540                 


Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val 
545                 550                 555                 560 


His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr 
                565                 570                 575     


Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly 
            580                 585                 590         


Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile 
        595                 600                 605             


Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val 
    610                 615                 620                 


Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser 
625                 630                 635                 640 


Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu 
                645                 650                 655     


Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro 
            660                 665                 670         


Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val 
        675                 680                 685             


Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met 
    690                 695                 700                 


His Glu Gly Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser 
705                 710                 715                 720 


Pro Gly Lys 
            


<210>  12
<211>  2169
<212>  DNA
<213>  Artificial

<220>
<223>  Nucleic Acid Sequence Encoding BS1 Antibody Chain 2 with 10H2 
       Light and Heavy Chains and Human IgG1 CH1, CH2, and CH3 Domains 
       with "Knobs" Substitution

<400>  12
atgagggtcc ccgctcagct cctggggctc ctgctgctct ggctcccagg tgcacgatgt       60

gccggatccc aggctgttgt gactcaggaa tctgcactca ccacatcacc tggtgaaaca      120

gtcacactca cttgtcgctc aagtactggg gctgttacaa ctagtaacta tgccaactgg      180

gtccaagaaa aaccagatca tttattcact ggtctaatag gtggtaccaa caaccgacct      240

ccaggtgttc ctgccagatt ctcaggctcc ctgattggag acaaggctgc cctcaccatc      300

acaggggcac agattgagga tgaggcaata tatttctgtg ctctatggta cagcaaccat      360

ttggtgttcg gtggaggaac caaactgact gtcctacgta cggtggctgc accatctgtc      420

ttcatcttcc cgccatctga tgagcagttg aaatctggaa ctgcctctgt tgtgtgcctg      480

ctgaataact tctatcccag agaggccaaa gtacagtgga aggtggataa cgccctccaa      540

tcgggtaact cccaggagag tgtcacagag caggacagca aggacagcac ctacagcctc      600

agcagcaccc tgacgctgag caaagcagac tacgagaaac acaaagtcta cgcctgcgaa      660

gtcacccatc agggcctgag ctcgcccgtc acaaagagct tcaacagggg tgagtgcggt      720

ggttctgggg gatctagcgg atcggggtct gggtcgactg gtacctcgtc aagcgggacg      780

ggtactagtg ctggtactac gggcacttca gctagcacct ctggctcggg gtccggcgag      840

gtgcagcttg ttgaaactgg tggaagattg gtgcagccta aagggtcatt gaaactctca      900

tgtgcagtct ctggaatcac cttcaagacc aatgccatga actgggtccg ccaggctcca      960

ggaaagggtt tggaatgggt tgctcgcata agaactaaaa gttataatta tgcaacatat     1020

tatgccgatt cagtgaaaga caggttcacc atctccagag atgattcaca aagcattctc     1080

tatctgcaaa tgaacaattt gaaaactgag gacacagcca tgtatcactg tgtgagagag     1140

ggccgctggg gccaagggac tctggtcact gtctctgcag ctagcaccaa gggcccatcg     1200

gtcttccccc tggcaccctc ctccaagagc acctctgggg gcacagcggc cctgggctgc     1260

ctggtcaagg actacttccc cgaaccggtg acggtgtcgt ggaactcagg cgccctgacc     1320

agcggcgtgc acaccttccc ggctgtccta cagtcctcag gactctactc cctcagcagc     1380

gtggtgaccg tgccctccag cagcttgggc acccagacct acatctgcaa cgtgaatcac     1440

aagcccagca acaccaaggt ggacaagaaa gttgagccca aatcttgtga caaaactcac     1500

acatgcccac cgtgcccagc acctgaactc ctggggggac cgtcagtctt cctcttcccc     1560

ccaaaaccca aggacaccct catgatctcc cggacccctg aggtcacatg cgtggtggtg     1620

gacgtgagcc acgaagaccc tgaggtcaag ttcaactggt acgtggacgg cgtggaggtg     1680

cataatgcca agacaaagcc gcgggaggag cagtacaaca gcacgtaccg tgtggtcagc     1740

gtcctcaccg tcctgcacca ggactggctg aatggcaagg agtacaagtg caaggtctcc     1800

aacaaagccc tcccagcccc catcgagaaa accatctcca aagccaaagg gcagccccga     1860

gaaccacagg tgtacaccct gcccccatcc cgggaggaga tgaccaagaa ccaggtcagc     1920

ctgtggtgcc tggtcaaagg cttctatccc agcgacatcg ccgtggagtg ggagagcaat     1980

gggcagccgg agaacaacta caagaccacg cctcccgtgc tggactccga cggctccttc     2040

ttcctctaca gcaagctcac cgtggacaag agcaggtggc agcaggggaa cgtcttctca     2100

tgctccgtga tgcatgaggg tctgcacaac cactacacgc agaagagcct ctccctgtct     2160

ccgggtaaa                                                             2169


<210>  13
<211>  40
<212>  PRT
<213>  Artificial

<220>
<223>  Exemplary Linker

<400>  13

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


Ser Ser Ser Gly Thr Gly Thr Ser Ala Gly Thr Thr Gly Thr Ser Ala 
            20                  25                  30          


Ser Thr Ser Gly Ser Gly Ser Gly 
        35                  40  


<210>  14
<211>  467
<212>  PRT
<213>  Artificial

<220>
<223>  BS2 10H2 Heavy Chain

<400>  14

Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp Leu Pro 
1               5                   10                  15      


Gly Ala Arg Cys Ala Gly Ser Glu Val Gln Leu Val Glu Thr Gly Gly 
            20                  25                  30          


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


Gly Ile Thr Phe Lys Thr Asn Ala Met Asn Trp Val Arg Gln Ala Pro 
    50                  55                  60                  


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


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


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


Thr Glu Asp Thr Ala Met Tyr His Cys Val Arg Glu Gly Arg Trp Gly 
        115                 120                 125             


Gln Gly Thr Leu Val Thr Val Ser Ala Ala Ser Thr Lys Gly Pro Ser 
    130                 135                 140                 


Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala 
145                 150                 155                 160 


Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val 
                165                 170                 175     


Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala 
            180                 185                 190         


Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val 
        195                 200                 205             


Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His 
    210                 215                 220                 


Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys 
225                 230                 235                 240 


Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly 
                245                 250                 255     


Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met 
            260                 265                 270         


Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His 
        275                 280                 285             


Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val 
    290                 295                 300                 


His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr 
305                 310                 315                 320 


Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly 
                325                 330                 335     


Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile 
            340                 345                 350         


Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val 
        355                 360                 365             


Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser 
    370                 375                 380                 


Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu 
385                 390                 395                 400 


Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro 
                405                 410                 415     


Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val 
            420                 425                 430         


Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met 
        435                 440                 445             


His Glu Gly Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser 
    450                 455                 460                 


Pro Gly Lys 
465         


<210>  15
<211>  490
<212>  PRT
<213>  Artificial

<220>
<223>  BS2 10H2+Tocilizumab scFv Light Chain

<400>  15

Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp Leu Pro 
1               5                   10                  15      


Gly Ala Arg Cys Ala Gly Ser Gln Ala Val Val Thr Gln Glu Ser Ala 
            20                  25                  30          


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


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


Pro Asp His Leu Phe Thr Gly Leu Ile Gly Gly Thr Asn Asn Arg Pro 
65                  70                  75                  80  


Pro Gly Val Pro Ala Arg Phe Ser Gly Ser Leu Ile Gly Asp Lys Ala 
                85                  90                  95      


Ala Leu Thr Ile Thr Gly Ala Gln Ile Glu Asp Glu Ala Ile Tyr Phe 
            100                 105                 110         


Cys Ala Leu Trp Tyr Ser Asn His Leu Val Phe Gly Gly Gly Thr Lys 
        115                 120                 125             


Leu Thr Val Leu Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro 
    130                 135                 140                 


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


Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp 
                165                 170                 175     


Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp 
            180                 185                 190         


Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys 
        195                 200                 205             


Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln 
    210                 215                 220                 


Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys Gly 
225                 230                 235                 240 


Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Gln Glu Ser 
                245                 250                 255     


Gly Pro Gly Leu Val Arg Pro Ser Gln Thr Leu Ser Leu Thr Cys Thr 
            260                 265                 270         


Val Ser Gly Tyr Ser Ile Thr Ser Asp His Ala Trp Ser Trp Val Arg 
        275                 280                 285             


Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile Gly Tyr Ile Ser Tyr Ser 
    290                 295                 300                 


Gly Ile Thr Thr Tyr Asn Pro Ser Leu Lys Ser Arg Val Thr Met Leu 
305                 310                 315                 320 


Arg Asp Thr Ser Lys Asn Gln Phe Ser Leu Arg Leu Ser Ser Val Thr 
                325                 330                 335     


Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala Arg Ser Leu Ala Arg Thr 
            340                 345                 350         


Thr Ala Met Asp Tyr Trp Gly Gln Gly Ser Leu Val Thr Val Ser Ser 
        355                 360                 365             


Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp 
    370                 375                 380                 


Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 
385                 390                 395                 400 


Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser Ser Tyr Leu 
                405                 410                 415     


Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 
            420                 425                 430         


Tyr Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 
        435                 440                 445             


Gly Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Leu Gln Pro Glu 
    450                 455                 460                 


Asp Ile Ala Thr Tyr Tyr Cys Gln Gln Gly Asn Thr Leu Pro Tyr Thr 
465                 470                 475                 480 


Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 
                485                 490 


<210>  16
<211>  6
<212>  PRT
<213>  Artificial

<220>
<223>  Tocilizumab Heavy Chain Variable Region CDR1

<400>  16

Ser Asp His Ala Trp Ser 
1               5       


<210>  17
<211>  16
<212>  PRT
<213>  Artificial

<220>
<223>  Tocilizumab Heavy Chain Variable Region CDR2

<400>  17

Tyr Ile Ser Tyr Ser Gly Ile Thr Thr Tyr Asn Pro Ser Leu Lys Ser 
1               5                   10                  15      


<210>  18
<211>  10
<212>  PRT
<213>  Artificial

<220>
<223>  Tocilizumab Heavy Chain Variable Region CDR3

<400>  18

Ser Leu Ala Arg Thr Thr Ala Met Asp Tyr 
1               5                   10  


<210>  19
<211>  11
<212>  PRT
<213>  Artificial

<220>
<223>  Tocilizumab Light Chain Variable Region CDR1

<400>  19

Arg Ala Ser Gln Asp Ile Ser Ser Tyr Leu Asn 
1               5                   10      


<210>  20
<211>  7
<212>  PRT
<213>  Artificial

<220>
<223>  Tocilizumab Light Chain Variable Region CDR2

<400>  20

Tyr Thr Ser Arg Leu His Ser 
1               5           


<210>  21
<211>  9
<212>  PRT
<213>  Artificial

<220>
<223>  Tocilizumab Light Chain Variable Region CDR3

<400>  21

Gln Gln Gly Asn Thr Leu Pro Tyr Thr 
1               5                   


<210>  22
<211>  8
<212>  PRT
<213>  Artificial

<220>
<223>  10H2 Heavy Chain Variable Region CDR1

<400>  22

Gly Ile Thr Phe Lys Thr Asn Ala 
1               5               


<210>  23
<211>  10
<212>  PRT
<213>  Artificial

<220>
<223>  10H2 Heavy Chain Variable Region CDR2

<400>  23

Ile Arg Thr Lys Ser Tyr Asn Tyr Ala Thr 
1               5                   10  


<210>  24
<211>  5
<212>  PRT
<213>  Artificial

<220>
<223>  10H2 Heavy Chain Variable Region CDR3

<400>  24

Val Arg Glu Gly Arg 
1               5   


<210>  25
<211>  9
<212>  PRT
<213>  Artificial

<220>
<223>  10H2 Light Chain Variable Region CDR1

<400>  25

Thr Gly Ala Val Thr Thr Ser Asn Tyr 
1               5                   


<210>  26
<211>  3
<212>  PRT
<213>  Artificial

<220>
<223>  10H2 Light Chain Variable Region CDR2

<400>  26

Gly Thr Asn 
1           


<210>  27
<211>  9
<212>  PRT
<213>  Artificial

<220>
<223>  10H2 Light Chain Variable Region CDR3

<400>  27

Ala Leu Trp Tyr Ser Asn His Leu Val 
1               5                   


<210>  28
<211>  21
<212>  DNA
<213>  Artificial

<220>
<223>  Forward HK2 Human

<400>  28
ccagttcatt cacatcatca g                                                 21


<210>  29
<211>  20
<212>  DNA
<213>  Artificial

<220>
<223>  Forward ?-Tubulin Human

<400>  29
aggagtccag atcggcaatg                                                   20


<210>  30
<211>  20
<212>  DNA
<213>  Artificial

<220>
<223>  Forward alpha-Tubulin Mouse

<400>  30
cacacaagct cactcaccct                                                   20


<210>  31
<211>  26
<212>  DNA
<213>  Artificial

<220>
<223>  Forward beta-Actin Human

<400>  31
atgagctgcc tgacggccag gtcatc                                            26


<210>  32
<211>  20
<212>  DNA
<213>  Artificial

<220>
<223>  Forward GAPDH Human

<400>  32
accacagtcc atgccatcac                                                   20


<210>  33
<211>  21
<212>  DNA
<213>  Artificial

<220>
<223>  Forward RPL13A Human

<400>  33
agcctcatct gcaatgtagg g                                                 21


<210>  34
<211>  20
<212>  DNA
<213>  Artificial

<220>
<223>  Forward GAPDH Mouse

<400>  34
accacagtcc atgccatcac                                                   20


<210>  35
<211>  21
<212>  DNA
<213>  Artificial

<220>
<223>  Forward RPL13A Human

<400>  35
agcctcatct gcaatgtagg g                                                 21


<210>  36
<211>  20
<212>  DNA
<213>  Artificial

<220>
<223>  Forward RPL13A Mouse

<400>  36
aggggttggt attcatccgc                                                   20


<210>  37
<211>  20
<212>  DNA
<213>  Artificial

<220>
<223>  Forward 18s Human

<400>  37
gaggatgagg tggaacgtgt                                                   20


<210>  38
<211>  20
<212>  DNA
<213>  Artificial

<220>
<223>  Forward 18s Mouse

<400>  38
cggcgacgac ccattcgaac                                                   20


<210>  39
<211>  20
<212>  DNA
<213>  Artificial

<220>
<223>  Reverse HK2 Human

<400>  39
cttacacgag gtcacatagc                                                   20


<210>  40
<211>  20
<212>  DNA
<213>  Artificial

<220>
<223>  Reverse alpha-Tubulin Human

<400>  40
gtccccacca ccaatggttt                                                   20


<210>  41
<211>  23
<212>  DNA
<213>  Artificial

<220>
<223>  Reverse beta-Actin Human

<400>  41
ctgttattag ggatgtgact cca                                               23


<210>  42
<211>  21
<212>  DNA
<213>  Artificial

<220>
<223>  Reverse alpha-Tubulin Mouse

<400>  42
ctccttaatg tcacgcacga t                                                 21


<210>  43
<211>  26
<212>  DNA
<213>  Artificial

<220>
<223>  Reverse beta-Actin Human

<400>  43
tggtaccacc agacagcact gtgttg                                            26


<210>  44
<211>  20
<212>  DNA
<213>  Artificial

<220>
<223>  Reverse GAPDH Human

<400>  44
gccttctcca tggtggtgaa                                                   20


<210>  45
<211>  21
<212>  DNA
<213>  Artificial

<220>
<223>  Reverse GAPDH Mouse

<400>  45
caccaccctg ttgctgtagc c                                                 21


<210>  46
<211>  23
<212>  DNA
<213>  Artificial

<220>
<223>  Reverse RPL13A Human

<400>  46
tcagactcct cggattcttc ttt                                               23


<210>  47
<211>  20
<212>  DNA
<213>  Artificial

<220>
<223>  Reverse RPL13A Mouse

<400>  47
atgcctgctg aggctttgtt                                                   20


<210>  48
<211>  20
<212>  DNA
<213>  Artificial

<220>
<223>  Reverse 18s Human

<400>  48
agaagtgacg cagccctcta                                                   20


<210>  49
<211>  22
<212>  DNA
<213>  Artificial

<220>
<223>  Reverse 18s Mouse

<400>  49
gaatcgaacc ctgattcccc gt                                                22


