U.S. patent application number 12/156063 was filed with the patent office on 2009-10-15 for human gm-csf antibodies. Invention is credited to Antony Burgess, Christoph Renner, Andrew Scott.
Application Number | 20090259027 12/156063 |
Document ID | / |
Family ID | 27734567 |
Filed Date | 2009-10-15 |
United States Patent Application | 20090259027 |
Kind Code | A1 |
Renner; Christoph ; et al. | October 15, 2009 |
Chimeric antibodies, as well as fusion proteins which comprise chimeric antibodies, are disclosed. The antibodies bind to GM-CSF, CD-30, and G250 antigen. The fusion proteins include biologically active portions of tumor necrosis factor, or full length tumor necrosis factor. Expression vectors adapted for production of the antibodies, as well as methods for manufacturing these, are also disclosed.
Inventors: | Renner; Christoph; (Homburg, DE) ; Burgess; Antony; (Parkville/Melbourne, AU) ; Scott; Andrew; (Parkville/Melbourne, AU) |
Correspondence Address: |
FULBRIGHT & JAWORSKI, LLP 666 FIFTH AVE NEW YORK NY 10103-3198 US |
Family ID: | 27734567 |
Appl. No.: | 12/156063 |
Filed: | May 29, 2008 |
Application Number | Filing Date | Patent Number | ||
---|---|---|---|---|
10365123 | Feb 12, 2003 | 7381801 | ||
12156063 | ||||
60355838 | Feb 13, 2002 | |||
Current U.S. Class: | 530/387.9 ; 435/320.1; 435/325; 435/358; 536/23.53 |
Current CPC Class: | C07K 16/2878 20130101; C07K 2317/92 20130101; C07K 14/525 20130101; C07K 2319/33 20130101; C07K 16/3038 20130101; C07K 16/30 20130101; C07K 2317/73 20130101; C07K 2317/24 20130101; C07K 16/243 20130101; C07K 2319/00 20130101; A61P 29/00 20180101; C07K 2319/30 20130101; C07K 2319/75 20130101; C07K 16/40 20130101; C07K 2317/56 20130101; C07K 2317/734 20130101; C07K 2317/732 20130101 |
Class at Publication: | 530/387.9 ; 536/23.53; 435/320.1; 435/325; 435/358 |
International Class: | C07K 16/00 20060101 C07K016/00; C07H 21/04 20060101 C07H021/04; C12N 15/63 20060101 C12N015/63; C12N 5/10 20060101 C12N005/10 |
Sequence CWU 1
1
56127DNAArtificial SequenceOligonucleotide primer 1atgaaatgca
gctgggtcat sttcttc 27225DNAArtificial SequenceOligonucleotide
primer 2atgggatgga gctratcats ytctt 25327DNAArtificial
SequenceOligonucleotide primer 3atgaagwtgt ggttaaactg ggttttt
27424DNAArtificial SequenceOligonucleotide primer 4atgractttg
wytcagcttg rttt 24528DNAArtificial SequenceOligonucleotide primer
5atggactcca ggctcaamag ttttcctt 28627DNAArtificial
SequenceOligonucleotide primer 6atggctgtcy trgsgctrct cttctgc
27726DNAArtificial SequenceOligonucleotide primer 7atggratgga
gckggrtctt tmtctt 26823DNAArtificial SequenceOligonucleotide primer
8atgagagtgc tgattctttt gtg 23930DNAArtificial
SequenceOligonucleotide primer 9atggmttggg tgtggamctt gctattcctg
301027DNAArtificial SequenceOligonucleotide primer 10atgggcagac
ttacattctc attcctg 271128DNAArtificial SequenceOligonucleotide
primer 11atggattttg ggctgatttt ttttattg 281227DNAArtificial
SequenceOligonucleotide primer 12atgatggtgt taagtcttct gtacctg
271330DNAArtificial SequenceOligonucleotide primer 13atgaagttgc
ctgttaggct gttggtgctg 301429DNAArtificial SequenceOligonucleotide
primer 14atggagwcag acacactcct gytatgggt 291530DNAArtificial
SequenceOligonucleotide primer 15atgagtgtgc tcactcaggt cctggsgttg
301633DNAArtificial SequenceOligonucleotide primer 16atgaggrccc
ctgctcagwt tyttggmwtc ttg 331730DNAArtificial
SequenceOligonucleotide primer 17atggatttwc aggtgcagat twtcagcttc
301827DNAArtificial SequenceOligonucleotide primer 18atgaggtkcy
ytgytsagyt yctgrgg 271931DNAArtificial SequenceOligonucleotide
primer 19atgggcwtca agatggagtc acakwyycwg g 312031DNAArtificial
SequenceOligonucleotide primer 20atgtggggay ctktttycmm tttttcaatt g
312125DNAArtificial SequenceOligonucleotide primer 21atggtrtccw
casctcagtt ccttg 252227DNAArtificial SequenceOligonucleotide primer
22atgtatatat gtttgttgtc tatttct 272328DNAArtificial
SequenceOligonucleotide primer 23atggaagccc cagctcagct tctcttcc
282427DNAArtificial SequenceOligonucleotide primer 24atgaagtttc
cttctcaact tctgctc 272517DNAArtificial SequenceOligonucleotide
primer 25tggatggtgg gaagatg 172618DNAArtificial
SequenceOligonucleotide primer 26ccagtggata gacagatg 1827456DNAMus
musculusmurine 19/2 heavy chain variable region 27atggagctga
tcatgctctt cctcctgtca ggaactgcag gcgtccactc 50tgaggtccag cttcagcagt
caggacctga actggtgaaa cctggggcct 100cagtgaagat atcctgcaag
gcttctggat acactttcac tgactacaac 150atacactggg tgaaacagag
ccatggaaag agccttgact ggattggata 200tattgctcct tacagtggtg
gtactggtta caaccaggag ttcaagaaca 250gggccacatt gactgtagac
aaatcctcca gcacagccta catggagctc 300cgcagtctga catctgatga
ctctgcagtc tattactgtg ctagacgaga 350ccgtttccct tattactttg
actactgggg ccaaggcacc cctctcacag 400tctcctcagc caaaacgaca
cccccatctg tctatccact ggcaagggcg 450aattcc 45628139PRTMus
musculusamino acid sequence for murine 19/2 heavy chain variable
region 28Met Glu Leu Ile Met Leu Phe Leu Leu Ser Gly Thr Ala Gly
Val His1 5 10 15Ser Glu Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Val
Lys Pro Gly 20 25 30Ala Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr
Thr Phe Thr Asp 35 40 45Tyr Asn Ile His Trp Val Lys Gln Ser His Gly
Lys Ser Leu Asp Trp 50 55 60Ile Gly Tyr Ile Ala Pro Tyr Ser Gly Gly
Thr Gly Tyr Asn Gln Glu65 70 75 80Phe Lys Asn Arg Ala Thr Leu Thr
Val Asp Lys Ser Ser Ser Thr Ala 85 90 95Tyr Met Glu Leu Arg Ser Leu
Thr Ser Asp Asp Ser Ala Val Tyr Tyr 100 105 110Cys Ala Arg Arg Asp
Arg Phe Pro Tyr Phe Asp Tyr Trp Gly Gln Gly 115 120 125Thr Thr Leu
Arg Val Ser Ser Val Ser Gly Ser 130 13529450DNAMus musculusmurine
19/2 light chain variable region 29atgggcttca agatggagtc acagatccag
gtctttgtat acatgttgct 50gtggttgtct ggtgttgatg gagacattgt gatgatccag
tctcaaaaat 100tcgtatccac atcagtagga gacagggtca atatcacctg
caaggccagt 150cagaatgtgg gaagtaatgt agcctggttg caacagaaac
ctggacaatc 200tcctaaaacg ctgatttact cggcatcgta ccggtccggt
cgagtccctg 250atcgcttcac aggcagtgga tctggaacag atttcattct
taccatcact 300actgtgcagt ctgaagactt ggcagaatat ttctgtcagc
aatttaacag 350gtctcctctc acgttcggtt ctgggaccaa gttggaactg
aaacgggctg 400atgctgcacc aactgtatcc atcttcccac catccagtaa
gggcgaattc 45030150PRTMus musculusamino acid sequence for murine
19/2 light chain variable region 30Met Gly Phe Lys Met Glu Ser Gln
Ile Gln Val Phe Val Tyr Met Leu1 5 10 15Leu Trp Leu Ser Gly Val Asp
Gly Asp Ile Val Met Ile Gln Ser Gln 20 25 30Lys Phe Val Ser Thr Ser
Val Gly Asp Arg Val Asn Ile Thr Cys Lys 35 40 45Ala Ser Gln Asn Val
Gly Ser Asn Val Ala Trp Leu Gln Gln Lys Pro 50 55 60Gly Gln Ser Pro
Lys Thr Leu Ile Tyr Ser Ala Ser Tyr Arg Ser Gly65 70 75 80Arg Val
Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Ile 85 90 95Leu
Thr Ile Thr Thr Val Gln Ser Glu Asp Leu Ala Glu Tyr Phe Cys 100 105
110Gln Gln Phe Asn Arg Ser Pro Leu Thr Phe Gly Ser Gly Thr Lys Leu
115 120 125Glu Leu Lys Arg Ala Asp Ala Ala Pro Thr Val Ser Ile Phe
Pro Pro 130 135 140Ser Ser Lys Gly Glu Phe145 1503141DNAArtificial
Sequenceprimer used for the construction of the chimeric 19/2 light
chain 31catgtttaaa cgccgccacc atgggcttca agatggagtc a
413238DNAArtificial Sequenceprimer used for the construction of the
chimeric 19/2 light chain 32agaggatcca ctcacgtttc agttccactt
ggtcccag 383343DNAArtificial Sequenceprimer used for the
construction of the chimeric 19/2 heavy chain 33catgtttaaa
cgccgccacc atggagctga tcatgctctt cct 433437DNAArtificial
Sequenceprimer used for the construction of the chimeric 19/2 heavy
chain 34agaggatcca ctcacctgag gagactctga gagtggt
37356159DNAArtificial SequencepREN 19/2 LC Neo Vector 35ctcgagagcg
ggcagtgagc gcaacgcaat taatgtgagt tagctcactc 50attaggcacc ccaggcttta
cactttatgc tcccggctcg tatgttgtgt 100ggagattgtg agcggataac
aatttcacac agaattcgtg aggctccggt 150gcccgtcagt gggcagagcg
cacatcgccc acagtccccg agaagttggg 200gggaggggtc ggcaattgaa
ccggtgccta gagaaggtgg cgcggggtaa 250actgggaaag tgatgtcgtg
tactggctcc gcctttttcc cgagggtggg 300ggagaaccgt atataagtgc
agtagtcgcc gtgaacgttc tttttcgcaa 350cgggtttgcc gccagaacac
aggtaagtgc cgtgtgtggt tcccgcgggc 400ctggcctctt tacgggttat
ggcccttgcg tgccttgaat tacttccacg 450cccctggctg cagtacgtga
ttcttgatcc cgagcttcgg gttggaagtg 500ggtgggagag ttcgaggcct
tgcgcttaag gagccccttc gcctcgtgct 550tgagttgagg cctggcctgg
gcgctggggc cgccgcgtgc gaatctggtg 600gcaccttcgc gcctgtctcg
ctgctttcga taagtctcta gccatttaaa 650atttttgatg acctgctgcg
acgctttttt tctggcaaga tagtcttgta 700aatgcgggcc aagatctgca
cactggtatt tcggtttttg gggccgcggg 750cggcgacggg gcccgtgcgt
cccagcgcac atgttcggcg aggcggggcc 800tgcgagcgcg gccaccgaga
atcggacggg ggtagtctca agctggccgg 850cctgctctgg tgcctggcct
cgcgccgccg tgtatcgccc cgccctgggc 900ggcaaggctg gcccggtcgg
caccagttgc gtgagcggaa agatggccgc 950ttcccggccc tgctgcaggg
agctcaaaat ggaggacgcg gcgctcggga 1000gagcgggcgg gtgagtcacc
cacacaaagg aaaagggcct ttccgtcctc 1050agccgtcgct tcatgtgact
ccacggagta ccgggcgccg tccaggcacc 1100tcgattagtt ctcgagcttt
tggagtacgt cgtctttagg ttggggggag 1150gggttttatg cgatggagtt
tccccacact gagtgggtgg agactgaagt 1200taggccagct tggcacttga
tgtaattctc cttggaattt gccctttttg 1250agtttggatc ttggttcatt
ctcaagcctc agacagtggt tcaaagtttt 1300tttcttccat ttcaggtgta
cgcgtctcgg gaagctttag tttaaacgcc 1350gccacc atg ggc ttc aag atg gag
tca cag atc cag gtc ttt 1392 Met Gly Phe Lys Met Glu Ser Gln Ile
Gln Val Phe 1 5 10gta tac atg ttg ctg tgg ttg tct ggt gtt gat gga
gac att 1434Val Tyr Met Leu Leu Trp Leu Ser Gly Val Asp Gly Asp Ile
15 20 25gtg atg atc cag tct caa aaa ttc gta tcc aca tca gta gga
1476Val Met Ile Gln Ser Gln Lys Phe Val Ser Thr Ser Val Gly 30 35
40gac agg gtc aat atc acc tgc aag gcc agt cag aat gtg gga 1518Asp
Arg Val Asn Ile Thr Cys Lys Ala Ser Gln Asn Val Gly 45 50agt aat
gta gcc tgg ttg caa cag aaa cct gga caa tct cct 1560Ser Asn Val Ala
Trp Leu Gln Gln Lys Pro Gly Gln Ser Pro55 60 65aaa acg ctg att tac
tcg gca tcg tac cgg tcc ggt cga gtc 1602Lys Thr Leu Ile Tyr Ser Ala
Ser Tyr Arg Ser Gly Arg Val 70 75 80cct gat cgc ttc aca ggc agt gga
tct gga aca gat ttc att 1644Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly
Thr Asp Phe Ile 85 90 95ctt acc atc act act gtg cag tct gaa gac ttg
gca gaa tat 1686Leu Thr Ile Thr Thr Val Gln Ser Glu Asp Leu Ala Glu
Tyr 100 105 110ttc tgt cag caa ttt aac agg tct cct ctc acg ttc ggt
tct 1728Phe Cys Gln Gln Phe Asn Arg Ser Pro Leu Thr Phe Gly Ser 115
120ggg acc aag ttg gaa ctg aaa cgt gagtggatcc atctgggata 1772Gly
Thr Lys Leu Glu Leu Lys Arg125 130agcatgctgt tttctgtctg tccctaacat
gccctgtgat tatgcgcaaa 1822caacacaccc aagggcagaa ctttgttact
taaacaccat cctgtttgct 1872tctttcctca gga act gtg gct gca cca tct
gtc ttc atc ttc 1915 Thr Val Ala Ala Pro Ser Val Phe Ile Phe 135
140ccg cca tct gat gag cag ttg aaa tct gga act gcc tct gtt 1957Pro
Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val 145 150 155gtg
tgc ctg ctg aat aac ttc tat ccc aga gag gcc aaa gta 1999Val Cys Leu
Leu Asn Asn Phe Tyr Pro Arg Gly Ala Lys Val 160 165 170cag tgg aag
gtg gat aac gcc ctc caa tcg ggt aac tcc cag 2041Gln Trp Lys Val Asp
Asn Ala Leu Gln Ser Gly Asn Ser Gln 175 180gag agt gtc aca gag cag
gac agc aag gac agc acc tac agc 2083Glu Ser Val Thr Glu Gln Asp Ser
Lys Asp Ser Thr Tyr Ser185 190 195ctc agc agc acc ctg acg ctg agc
aaa gca gac tac gag aaa 2125Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala
Asp Tyr Glu Lys 200 205 210cac aaa gtc tac gcc tgc gaa gtc acc cat
cag ggc ctg agc 2167His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly
Leu Ser 215 220 225tcg ccc gtc aca aag agc ttc aac agg gga gag tgt
tga 2206Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 230
235gctagaacta actaactaag ctagcaacgg tttccctcta gcgggatcaa
2256ttccgccccc cccccctaac gttactggcc gaagccgctt ggaataaggc
2306cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg
2356caatgtgagg gcccggaaac ctggccctgt cttcttgacg agcattccta
2406ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg
2456aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc
2506gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg
2556gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag
2606tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc
2656aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta
2706tgggatctga tctggggcct cggtgcacat gctttacgtg tgtttagtcg
2756aggttaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt
2806tgaaaaacac gataatacca tggttgaaca agatggattg cacgcaggtt
2856ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
2906acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
2956cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
3006aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
3056gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
3106gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
3156agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
3206ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
3256acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
3306agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
3356atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
3406gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
3456ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
3506attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
3556cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
3606acgagttctt ctgagtcgat cgacctggcg taatagcgaa gaggcccgca
3656ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg
3706ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
3756gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
3806cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
3856gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
3906aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
3956cggtttttcg cctttgacgt tggagtccac gttctttaat agtggactct
4006tgttccaaac tggaacaaca ctcaacccta tctcggtcta tttataaggg
4056attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
4106tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact
4156tttcggggaa atgtgcgcgg aacccctata tttgtttatt tttctaaata
4206cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
4256ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
4306ttattccctt ttttgcggca ttttgcctta ctgtttttgc tcacccagaa
4356acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
4406ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
4456ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
4506gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
4556acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
4606atattacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
4656atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
4706gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
4756ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
4806gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
4856tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
4906aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
4956tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
5006cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
5056acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
5106gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
5156ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
5206tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
5256gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatg
5306ttcttgagat cctttttttc tgcacgtaat ctgctgcttg caaacaaaaa
5356accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
5406tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
5456cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
5506gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
5556gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
5606aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
5656ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
5706aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
5756ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
5806ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
5856gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
5906aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
5956gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
6006ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
6056tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
6106cgcgcgttgg ccgattcatt aatgcaggta tcacgaggcc ctttcgtctt 6156cac
6159366579DNAArtificial SequencepREN 19/2 HC DHFR Vector
36ctcgagagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
50attaggcacc ccaggcttta cactttatgc tcccggctcg tatgttgtgt
100ggagattgtg agcggataac aatttcacac agaattcgtg aggctccggt
150gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
200gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa
250actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg
300ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa
350cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc
400ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccacg
450ggtgggagag ttcgaggcct tgcgcttaag gagccccttc gcctcgtgct
500tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc gaatctggtg
550gcaccttcgc gcctgtctcg ctgctttcga taagtctcta gccatttaaa
600atttttgatg acctgctgcg acgctttttt tctggcaaga tagtcttgta
650aatgcgggcc aagatctgca cactggtatt tcggtttttg gggccgcggg
700cggcgacggg gcccgtgcgt cccagcgcac atgttcggcg aggcggggcc
750tgcgagcgcg gccaccgaga atcggacggg ggtagtctca agctggccgg
800cctgctctgg tgcctggcct cgcgccgccg tgtatcgccc cgccctgggc
850ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa agatggccgc
900ttcccggccc tgctgcaggg agctcaaaat ggaggacgcg gcgctcggga
950gagcgggcgg gtgagtcacc cacacaaagg aaaagggcct ttccgtcctc
1000agccgtcgct tcatgtgact ccacggagta ccgggcgccg tccaggcacc
1050tcgattagtt ctcgagcttt tggagtacgt cgtctttagg ttggggggag
1100gggttttatg cgatggagtt tccccacact gagtgggtgg agactgaagt
1150taggccagct tggcacttga tgtaattctc cttggaattt gccctttttg
1200agtttggatc ttggttcatt ctcaagcctc agacagtggt tcaaagtttt
1250tttcttccat ttcaggtgta cgcgtctcgg gaagctttag tttaaacgcc
1300gccacc atg gag ctg atc atg ctc ttc ctc ctg tca gga act 1342 Met
Glu Leu Ile Met Leu Phe Leu Leu Ser Gly Thr 1 5 10gca ggc gtc cac
tct gag gtc cag ctt cag cag tca gga cct 1384Ala Gly Val His Ser Glu
Val Gln Leu Gln Gln Ser Gly Pro 15 20 25gaa ctg gtg aaa cct ggg gcc
tca gtg aag ata tcc tgc aag 1426Glu Leu Val Lys Pro Gly Ala Ser Val
Lys Ile Ser Cys Lys 30 35 40gct tct gga tac act ttc act gac tac aac
ata cac tgg gtg 1468Ala Ser Gly Tyr Thr Phe Thr Asp Tyr Asn Ile His
Trp Val 45 50aaa cag agc cat gga aag agc ctt gac tgg att gga tat
att 1510Lys Gln Ser His Gly Lys Ser Leu Asp Trp Ile Gly Tyr Ile55
60 65gct cct tac agt ggt ggt act ggt tac aac cag gag ttc aag
1552Ala Pro Tyr Ser Gly Gly Thr Gly Tyr Asn Gln Glu Phe Lys 70 75
80aac agg gcc aca ttg act gta gac aaa tcc tcc agc aca gcc 1594Asn
Arg Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala 85 90 95tac atg
gag ctc cgc agt ctg aca tct gat gac tct gca gtc 1636Tyr Met Glu Leu
Arg Ser Leu Thr Ser Asp Asp Ser Ala Val 100 105 110tat tac tgt gct
aga cga gac cgt ttc cct tat tac ttt gac 1678Tyr Tyr Cys Ala Arg Arg
Asp Arg Phe Pro Tyr Tyr Phe Asp 115 120tac tgg ggc caa ggc acc act
ctc aga gtc tcc tca gtgagt 1720Tyr Trp Gly Gln Gly Thr Thr Leu Arg
Val Ser Ser125 130 135ggatcctctg cgcctgggcc cagctctgtc ccacaccgcg
gtcacatggc 1770accacctctc ttgcagcc tcc acc aag ggc cca tcg gtc ttc
1812 Ser Thr Lys Gly Pro Ser Val Phe 140ccc ctg gca ccc tcc tcc aag
agc acc tct ggg ggc aca 1851Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser
Gly Gly Thr145 150 155gcg gcc ctg ggc tgc ctg gtc aag gac tac ttc
ccc gaa ccg 1893Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu
Pro 160 165 170gtg acg gtg tcg tgg aac tca ggc gcc ctg acc agc ggc
gtg 1935Val Thr Val Ser Trp Asn Ser Gly Ala Lys Thr Ser Gly Val 175
180 185cac acc ttc ccg gct gtc cta cag tcc tca gga ctc tac tcc
1977His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser 190
195ctc agc agc gtg gtg acc gtg ccc tcc agc agc ttg ggc acc 2019Leu
Ser Ser Val Val Ser Val Pro Ser Ser Ser Leu Gly Thr200 205 210cag
acc tac atc tgc aac gtg aat cac aag ccc agc aac acc 2061Gln Thr Tyr
Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr 215 220 225aag gtg gac
aag aaa gtt gag ccc aaa tct tgt gac aaa act 2103Lys Val Asn Lys Lys
Val Glu Pro Lys Ser Cys Asn Lys Thr 230 235 240cac aca tgc cca ccg
tgc cca gca cct gaa ctc ctg ggg gga 2145His Thr Cys Pro Pro Cys Pro
Ala Pro Glu Leu Leu Gly Gly 245 250 255ccg tca gtc ttc ctc ttc ccc
cca aaa ccc aag gac acc ctc 2187Pro Ser Val Phe Leu Phe Pro Pro Lys
Pro Lys Asn Thr Leu 260 265atg atc tcc cgg acc cct gag gtc aca tgc
gtg gtg gtg gac 2229Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val
Val Asn270 275 280gtg agc cac gaa gac cct gag gtc aag ttc aac tgg
tac gtg 2271Val Ser His Glu Asn Pro Glu Val Lys Phe Asn Trp Tyr Val
285 290 295gac ggc gtg gag gtg cat aac gcc aag aca aag ccg cgg gag
2313Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 300 305
310gag cag tac aac agc acg tac cgg gtg gtc agc gtc ctc acc 2355Glu
Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr 315 320 325gtc
ctg cac cag gac tgg ctg aat ggc aag gag tac aag tgc 2397Val Leu His
Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys 330 335aag gtc tcc aac
aaa gcc ctc cca gcc ccc atc gag aaa acc 2439Lys Val Ser Asn Lys Ala
Leu Pro Ala Pro Ile Glu Lys Thr340 345 350atc tcc aaa gcc aaa ggg
cag ccc cga gaa cca cag gtg tac 2481Ile Ser Lys Ala Lys Gly Gln Pro
Arg Glu Pro Gln Val Tyr 355 360 365acc ctg ccc cca tcc cgg gag gag
atg acc aag aac cag gtc 2523Thr Leu Pro Pro Ser Arg Glu Glu Met Thr
Lys Asn Gln Val 370 375 380agc ctg acc tgc ctg gtc aaa ggc ttc tat
ccc agc gac atc 2565Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser
Asn Ile 385 390 395gcc gtg gag tgg gag agc aat ggg cag ccg gag aac
aac tac 2607Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr
400 405aag acc acg cct ccc gtg ctg gac tcc gac ggc tcc ttc ttc
2649Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe410 415
420ctc tac agc aag ctc acc gtg gac aag agc agg tgg cag cag 2691Leu
Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln 425 430 435ggg
aac gtc ttc tca tgc tcc gtg atg cat gag gct ctg cac 2733Gly Asn Val
Phe Ser Cys Ser Val Met His Glu Ala Leu His 440 445 450aac cac tac
acg cag aag agc ctc tcc ctg tct ccg ggt aaa 2775Asn His Tyr Thr Gln
Lys Ser Leu Ser Leu Ser Pro Gly Lys 455 460 465tga gctagaaact
aactaagcta gcaacggttt ccctctagcg ggatcaattc 2828cgcccccccc
ccctaacgtt actggccgaa gccgcttgga ataaggccgg 2878tgtgcgtttg
tctatatgtt attttccacc atattgccgt cttttggcaa 2928tgtgagggcc
cggaaacctg gccctgtctt cttgacgagc attcctaggg 2978gtctttcccc
tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag 3028gaagcagttc
ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac 3078cctttgcagg
cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc 3128aaaagccacg
tgtataagat acacctgcaa aggcggcaca accccagtgc 3178cacgttgtga
gttggatagt tgtggaaaga gtcaaatggc tctcctcaag 3228cgtattcaac
aaggggctga aggatgccca gaaggtaccc cattgtatgg 3278gatctgatct
ggggcctcgg tgcacatgct ttacgtgtgt ttagtcgagg 3328ttaaaaaacg
tctaggcccc ccgaaccacg gggacgtggt tttcctttga 3378aaaacacgat
aataccatgg ttcgaccatt gaactgcatc gtcgccgtgt 3428cccaaaatat
ggggattggc aagaacggag acctaccctg gcctccgctc 3478aggaacgagt
tcaagtactt ccaaagaatg accacaacct cttcagtgga 3528aggtaaacag
aatctggtga ttatgggtag gaaaacctgg ttctccattc 3578ctgagaagaa
tcgaccttta aaggacagaa ttaatggttc gatatagttc 3628tcagtagaga
actcaaagaa ccaccacgag gagctcattt tcttgccaaa 3678agtttggatg
atgccttaag acttattgaa caaccggaat tggcaagtaa 3728agtagacatg
gtttggatag tcggaggcag ttctgtttac caggaagcca 3778tgaatcaacc
aggccacctc agactctttg tgacaaggat catgcaggaa 3828tttgaaagtg
acacgttttt cccagaaatt gatttgggga aatataaact 3878tctcccagaa
tacccaggcg tcctctctga ggtccaggag gaaaaaggca 3928tcaagtataa
gtttgaagtc tacgagaaga aagactaaca ggaagatgct 3978ttcaagttct
ctgctcccct cctaaagcta tgcattttta taagaccatg 4028ggacttttgc
tggtcgatcg acctggcgta atagcgaaga ggcccgcacc 4078gatcgccctt
cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc 4128ctgtagcggc
gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga 4178ccgctacact
tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct 4228tcctttctcg
ccacgttcgc cggctttccc cgtcaagctc taaatcgggg 4278gctcccttta
gggttccgat ttagtgcttt acggcacctc gaccccaaaa 4328aacttgatta
gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg 4378gtttttcgcc
tttgacgttg gagtccacgt tctttaatag tggactcttg 4428ttccaaactg
gaacaacact caaccctatc tcggtctatt tataagggat 4478tttgccgatt
tcggcctatt ggttaaaaaa tgagctgatt taacaaaatt 4528taacgcgaat
tttaacaaaa tattaacgct tacaatttag gtggcacttt 4578tcggggaaat
gtgcgcggaa cccctatatt tgtttatttt tctaaataca 4628ttcaaatatg
tatccgctca tgagacaata accctgataa atgcttcaat 4678aatattgaaa
aaggaagagt atgagtattc aacatttccg tgtcgccctt 4728attccctttt
ttgcggcatt ttgccttact gtttttgctc acccagaaac 4778gctggtgaaa
gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt 4828acatcgaact
ggatctcaac agcggtaaga tccttgagag ttttcgcccc 4878gaagaacgtt
ttccaatgat gagcactttt aaagttctgc tatgtggcgc 4928ggtattatcc
cgtattgacg ccgggcaaga gcaactcggt cgccgcatac 4978actattctca
gaatgacttg gttgagtact caccagtcac agaaaagcat 5028attacggatg
gcatgacagt aagagaatta tgcagtgctg ccataaccat 5078gagtgataac
actgcggcca acttacttct gacaacgatc ggaggaccga 5128aggagctaac
cgcttttttg cacaacatgg gggatcatgt aactcgcctt 5178gatcgttggg
aaccggagct gaatgaagcc ataccaaacg acgagcgtga 5228caccacgatg
cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg 5278gcgaactact
tactctagct tcccggcaac aattaataga ctggatggag 5328gcggataaag
ttgcaggacc acttctgcgc tcggcccttc cggctggctg 5378gtttattgct
gataaatctg gagccggtga gcgtgggtct cgcggtatca 5428ttgcagcact
ggggccagat ggtaagccct cccgtatcgt agttatctac 5478acgacgggga
gtcaggcaac tatggatgaa cgaaatagac agatcgctga 5528gataggtgcc
tcactgatta agcattggta actgtcagac caagtttact 5578catatatact
ttagattgat ttaaaacttc atttttaatt taaaaggatc 5628taggtgaaga
tcctttttga taatctcatg accaaaatcc cttaacgtga 5678gttttcgttc
cactgagcgt cagaccccgt agaaaagatc aaaggatgtt 5728cttgagatcc
tttttttctg cacgtaatct gctgcttgca aacaaaaaac 5778caccgctacc
agcggtggtt tgtttgccgg atcaagagct accaactctt 5828tttccgaagg
taactggctt cagcagagcg cagataccaa atactgtcct 5878tctagtgtag
ccgtagttag gccaccactt caagaactct gtagcaccgc 5928ctacatacct
cgctctgcta atcctgttac cagtggctgc tgccagtggc 5978gataagtcgt
gtcttaccgg gttggactca agacgatagt taccggataa 6028ggcgcagcgg
tcgggctgaa cggggggttc gtgcacacag cccagcttgg 6078agcgaacgac
ctacaccgaa ctgagatacc tacagcgtga gctatgagaa 6128agcgccacgc
ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg 6178cagggtcgga
acaggagagc gcacgaggga gcttccaggg ggaaacgcct 6228ggtatcttta
tagtcctgtc gggtttcgcc acctctgact tgagcgtcga 6278tttttgtgat
gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa 6328cgcggccttt
ttacggttcc tggccttttg ctggcctttt gctcacatgt 6378tctttcctgc
gttatcccct gattctgtgg ataaccgtat taccgccttt 6428gagtgagctg
ataccgctcg ccgcagccga acgaccgagc gcagcgagtc 6478agtgagcgag
gaagcggaag agcgcccaat acgcaaaccg cctctccccg 6528cgcgttggcc
gattcattaa tgcaggtatc acgaggccct ttcgtcttca 6578c
65793724DNAArtificial Sequenceprimer 37ttcttgaagt ctggtgatgc tgcc
243831DNAArtificial Sequenceprimer 38caagctagcc ctctaagact
cctcccctgt t 313933DNAArtificial Sequenceprimer 39gaactcgagt
catttacccg gagacaggga gag 334032DNAArtificial Sequenceprimer
40gcgccatggc ccaggtgcaa ctgcagcagt ca 324134DNAArtificial
Sequenceprimer 41cagggatcca ctcacctgag gagacggtga ccgt
344233DNAArtificial Sequenceprimer 42agcgccatgg acatcgagct
cactcagtct cca 334335DNAArtificial Sequenceprimer 43cagggatcca
actcacgttt gatttccagc ttggt 3544465DNAMus Musculusnucleotide
sequence of murine heavy chain variable region 44gtttaaacgc
cgccaccatg aactggacct ggaccgtgtt ttgcctgctc 50gctgtggctc ctggggccca
cagcgccatg gcccaggtgc aactgcagca 100gtcaggggct gagctggcta
gacctggggc ttcagtgaag atgtcctgca 150aggcttctgg ctacaccttt
actacctaca caatacactg ggtaagacag 200aggcctggac acgatctgga
atggattgga tacattaatc ctagcagtgg 250atattctgac tacaatcaaa
gcttcaaggg caagaccaca ttgactgcag 300acaagtcctc caacacagcc
tacatgcaac tgaacagcct gacatctgag 350gactctgcgg tctattactg
tgcaagaaga gcggactatg gtaactacga 400atatacctgg tttgcttact
ggggccaagg gaccacggtc accgtctcct 450caggtgagtg gatcc 46545145PRTMus
musculusamino acid sequence for murine heavy chain variable region
45Met Asn Trp Thr Trp Thr Val Phe Cys Leu Leu Ala Val Ala Pro Gly1
5 10 15Ala His Ser Ala Met Ala Gln Val Gln Leu Gln Gln Ser Gly Ala
Glu 20 25 30Leu Ala Arg Pro Gly Ala Ser Val Lys Met Ser Cys Lys Ala
Ser Gly 35 40 45Tyr Thr Phe Thr Thr Tyr Thr Ile His Trp Val Arg Gln
Arg Pro Gly 50 55 60His Asp Leu Glu Trp Ile Gly Tyr Ile Asn Pro Ser
Ser Gly Tyr Ser65 70 75 80Asp Tyr Asn Gln Ser Phe Lys Gly Lys Thr
Thr Leu Thr Ala Asp Lys 85 90 95Ser Ser Asn Thr Ala Tyr Met Gln Leu
Asn Ser Leu Thr Ser Glu Asp 100 105 110Ser Ala Val Tyr Tyr Cys Ala
Arg Arg Ala Asp Tyr Gly Asn Tyr Glu 115 120 125Tyr Thr Trp Phe Ala
Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser 130 135
140Ser14546415DNAMus musculusnucleotide sequence for murine light
chain variable region 46gtttaaacgc cgccaccatg aactggacct ggaccgtgtt
ttgcctgctc 50gctgtggctc ctggggccca cagcgccatg gacatcgagc tcactcagtc
100tccaaaattc atgtccacat cagtaggaga cagggtcaac gtcacctaca
150aggccagtca gaatgtgggt actaatgtag cctggtttca acaaaaacca
200gggcaatctc ctaaagttct gatttactcg gcatcttacc gatacagtgg
250agtccctgat cgcttcacag gcagtggatc tggaacagat ttcactctca
300ccatcagcaa tgtgcagtct gaagacttgg cagagtattt ctgtcagcaa
350tatcacacct atcctctcac gttcggaggg ggcaccaagc tggaaatcaa
400acgtgagttg gatcc 41547129PRTMus musculusamino acid sequence for
murine light chain variable region 47Met Asn Trp Thr Trp Thr Val
Phe Cys Leu Leu Ala Val Ala Pro Gly1 5 10 15Ala His Ser Ala Met Asp
Ile Glu Leu Thr Gln Ser Pro Lys Phe Met 20 25 30Ser Thr Ser Val Gly
Asp Arg Val Asn Val Thr Tyr Lys Ala Ser Gln 35 40 45Asn Val Gly Thr
Asn Val Ala
Trp Phe Gln Gln Lys Pro Gly Gln Ser 50 55 60Pro Lys Val Leu Ile Tyr
Ser Ala Ser Tyr Arg Tyr Ser Gly Val Pro65 70 75 80Asp Arg Phe Thr
Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile 85 90 95Ser Asn Val
Gln Ser Glu Asp Leu Ala Glu Tyr Phe Cys Gln Gln Tyr 100 105 110His
Thr Tyr Pro Leu Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 115 120
125Arg 485759DNAArtificial SequenceLight chain expression vector
pREN-Neo 48ctcgagagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
50attaggcacc ccaggcttta cactttatgc tcccggctcg tatgttgtgt
100ggagattgtg agcggataac aatttcacac agaattcgtg aggctccggt
150gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
200gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa
250actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg
300ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa
350cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc
400ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccacg
450cccctggctg cagtacgtga ttcttgatcc cgagcttcgg gttggaagtg
500ggtgggagag ttcgaggcct tgcgcttaag gagccccttc gcctcgtgct
550tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc gaatctggtg
600gcaccttcgc gcctgtctcg ctgctttcga taagtctcta gccatttaaa
650atttttgatg acctgctgcg acgctttttt tctggcaaga tagtcttgta
700aatgcgggcc aagatctgca cactggtatt tcggtttttg gggccgcggg
750cggcgacggg gcccgtgcgt cccagcgcac atgttcggcg aggcggggcc
800tgcgagcgcg gccaccgaga atcggacggg ggtagtctca agctggccgg
850cctgctctgg tgcctggcct cgcgccgccg tgtatcgccc cgccctgggc
900ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa agatggccgc
950ttcccggccc tgctgcaggg agctcaaaat ggaggacgcg gcgctcggga
1000gagcgggcgg gtgagtcacc cacacaaagg aaaagggcct ttccgtcctc
1050agccgtcgct tcatgtgact ccacggagta ccgggcgccg tccaggcacc
1100tcgattagtt ctcgagcttt tggagtacgt cgtctttagg ttggggggag
1150gggttttatg cgatggagtt tccccacact gagtgggtgg agactgaagt
1200taggccagct tggcacttga tgtaattctc cttggaattt gccctttttg
1250agtttggatc ttggttcatt ctcaagcctc agacagtggt tcaaagtttt
1300tttcttccat ttcaggtgta cgcgtctcgg gaagctttag tttaaacgcc
1350gtgagtggat ccatctggga taagcatgct gttttctgtc tgtccctaac
1400atgccctgtg attatgcgca aacaacacac ccaagggcag aactttgtta
1450cttaaacacc atcctgtttg cttctttcct cagga act gtg gct gca cca 1500
Thr Val Ala Ala Pro 1 5tct gtc ttc atc ttc ccg cca tct gat gag cag
ttg aaa tct gga 1545Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu
Lys Ser Gly 10 15 20act gcc tct gtt gtg tgc ctg ctg aat aac ttc tat
ccc aga gag 1590Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro
Arg Glu 25 30 35gcc aaa gta cag tgg aag gtg gat aac gcc ctc caa tcg
ggt aac 1635Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly
Asn 40 45 50tcc cag gag agt gtc aca gag cag gac agc aag gac agc acc
tac 1680Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr
55 60 65agc ctc agc agc acc ctg acg ctg agc aaa gca gac tac gag aaa
1725Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys 70
75 80cac aaa gtc tac gcc tgc gaa gtc acc cat cag ggc ctg agc tcg
1770His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser 85
90 95ccc gtc aca aag agc ttc aac agg gga gag tgt tga 1806Pro Val
Thr Lys Ser Phe Asn Arg Gly Glu Cys 100 105gctagaacta actaactaag
ctagcaacgg tttccctcta gcgggatcaa 1856ttccgccccc cccccctaac
gttactggcc gaagccgctt ggaataaggc 1906cggtgtgcgt ttgtctatat
gttattttcc accatattgc cgtcttttgg 1956caatgtgagg gcccggaaac
ctggccctgt cttcttgacg agcattccta 2006ggggtctttc ccctctcgcc
aaaggaatgc aaggtctgtt gaatgtcgtg 2056aaggaagcag ttcctctgga
agcttcttga agacaaacaa cgtctgtagc 2106gaccctttgc aggcagcgga
accccccacc tggcgacagg tgcctctgcg 2156gccaaaagcc acgtgtataa
gatacacctg caaaggcggc acaaccccag 2206tgccacgttg tgagttggat
agttgtggaa agagtcaaat ggctctcctc 2256aagcgtattc aacaaggggc
tgaaggatgc ccagaaggta ccccattgta 2306tgggatctga tctggggcct
cggtgcacat gctttacgtg tgtttagtcg 2356aggttaaaaa acgtctaggc
cccccgaacc acggggacgt ggttttcctt 2406tgaaaaacac gataatacca
tggttgaaca agatggattg cacgcaggtt 2456ctccggccgc ttgggtggag
aggctattcg gctatgactg ggcacaacag 2506acaatcggct gctctgatgc
cgccgtgttc cggctgtcag cgcaggggcg 2556cccggttctt tttgtcaaga
ccgacctgtc cggtgccctg aatgaactgc 2606aggacgaggc agcgcggcta
tcgtggctgg ccacgacggg cgttccttgc 2656gcagctgtgc tcgacgttgt
cactgaagcg ggaagggact ggctgctatt 2706gggcgaagtg ccggggcagg
atctcctgtc atctcacctt gctcctgccg 2756agaaagtatc catcatggct
gatgcaatgc ggcggctgca tacgcttgat 2806ccggctacct gcccattcga
ccaccaagcg aaacatcgca tcgagcgagc 2856acgtactcgg atggaagccg
gtcttgtcga tcaggatgat ctggacgaag 2906agcatcaggg gctcgcgcca
gccgaactgt tcgccaggct caaggcgcgc 2956atgcccgacg gcgaggatct
cgtcgtgacc catggcgatg cctgcttgcc 3006gaatatcatg gtggaaaatg
gccgcttttc tggattcatc gactgtggcc 3056ggctgggtgt ggcggaccgc
tatcaggaca tagcgttggc tacccgtgat 3106attgctgaag agcttggcgg
cgaatgggct gaccgcttcc tcgtgcttta 3156cggtatcgcc gctcccgatt
cgcagcgcat cgccttctat cgccttcttg 3206acgagttctt ctgagtcgat
cgacctggcg taatagcgaa gaggcccgca 3256ccgatcgccc ttcccaacag
ttgcgcagcc tgaatggcga atgggacgcg 3306ccctgtagcg gcgcattaag
cgcggcgggt gtggtggtta cgcgcagcgt 3356gaccgctaca cttgccagcg
ccctagcgcc cgctcctttc gctttcttcc 3406cttcctttct cgccacgttc
gccggctttc cccgtcaagc tctaaatcgg 3456gggctccctt tagggttccg
atttagtgct ttacggcacc tcgaccccaa 3506aaaacttgat tagggtgatg
gttcacgtag tgggccatcg ccctgataga 3556cggtttttcg cctttgacgt
tggagtccac gttctttaat agtggactct 3606tgttccaaac tggaacaaca
ctcaacccta tctcggtcta tttataaggg 3656attttgccga tttcggccta
ttggttaaaa aatgagctga tttaacaaaa 3706tttaacgcga attttaacaa
aatattaacg cttacaattt aggtggcact 3756tttcggggaa atgtgcgcgg
aacccctata tttgtttatt tttctaaata 3806cattcaaata tgtatccgct
catgagacaa taaccctgat aaatgcttca 3856ataatattga aaaaggaaga
gtatgagtat tcaacatttc cgtgtcgccc 3906ttattccctt ttttgcggca
ttttgcctta ctgtttttgc tcacccagaa 3956acgctggtga aagtaaaaga
tgctgaagat cagttgggtg cacgagtggg 4006ttacatcgaa ctggatctca
acagcggtaa gatccttgag agttttcgcc 4056ccgaagaacg ttttccaatg
atgagcactt ttaaagttct gctatgtggc 4106gcggtattat cccgtattga
cgccgggcaa gagcaactcg gtcgccgcat 4156acactattct cagaatgact
tggttgagta ctcaccagtc acagaaaagc 4206atattacgga tggcatgaca
gtaagagaat tatgcagtgc tgccataacc 4256atgagtgata acactgcggc
caacttactt ctgacaacga tcggaggacc 4306gaaggagcta accgcttttt
tgcacaacat gggggatcat gtaactcgcc 4356ttgatcgttg ggaaccggag
ctgaatgaag ccataccaaa cgacgagcgt 4406gacaccacga tgcctgtagc
aatggcaaca acgttgcgca aactattaac 4456tggcgaacta cttactctag
cttcccggca acaattaata gactggatgg 4506aggcggataa agttgcagga
ccacttctgc gctcggccct tccggctggc 4556tggtttattg ctgataaatc
tggagccggt gagcgtgggt ctcgcggtat 4606cattgcagca ctggggccag
atggtaagcc ctcccgtatc gtagttatct 4656acacgacggg gagtcaggca
actatggatg aacgaaatag acagatcgct 4706gagataggtg cctcactgat
taagcattgg taactgtcag accaagttta 4756ctcatatata ctttagattg
atttaaaact tcatttttaa tttaaaagga 4806tctaggtgaa gatccttttt
gataatctca tgaccaaaat cccttaacgt 4856gagttttcgt tccactgagc
gtcagacccc gtagaaaaga tcaaaggatg 4906ttcttgagat cctttttttc
tgcacgtaat ctgctgcttg caaacaaaaa 4956accaccgcta ccagcggtgg
tttgtttgcc ggatcaagag ctaccaactc 5006tttttccgaa ggtaactggc
ttcagcagag cgcagatacc aaatactgtc 5056cttctagtgt agccgtagtt
aggccaccac ttcaagaact ctgtagcacc 5106gcctacatac ctcgctctgc
taatcctgtt accagtggct gctgccagtg 5156gcgataagtc gtgtcttacc
gggttggact caagacgata gttaccggat 5206aaggcgcagc ggtcgggctg
aacggggggt tcgtgcacac agcccagctt 5256ggagcgaacg acctacaccg
aactgagata cctacagcgt gagctatgag 5306aaagcgccac gcttcccgaa
gggagaaagg cggacaggta tccggtaagc 5356ggcagggtcg gaacaggaga
gcgcacgagg gagcttccag ggggaaacgc 5406ctggtatctt tatagtcctg
tcgggtttcg ccacctctga cttgagcgtc 5456gatttttgtg atgctcgtca
ggggggcgga gcctatggaa aaacgccagc 5506aacgcggcct ttttacggtt
cctggccttt tgctggcctt ttgctcacat 5556gttctttcct gcgttatccc
ctgattctgt ggataaccgt attaccgcct 5606ttgagtgagc tgataccgct
cgccgcagcc gaacgaccga gcgcagcgag 5656tcagtgagcg aggaagcgga
agagcgccca atacgcaaac cgcctctccc 5706cgcgcgttgg ccgattcatt
aatgcaggta tcacgaggcc ctttcgtctt 5756cac 5759496207DNAArtificial
SequenceHeavy chain expression vector pREN-DHFR 49ctcgagagcg
ggcagtgagc gcaacgcaat taatgtgagt tagctcactc 50attaggcacc ccaggcttta
cactttatgc tcccggctcg tatgttgtgt 100ggagattgtg agcggataac
aatttcacac agaattcgtg aggctccggt 150gcccgtcagt gggcagagcg
cacatcgccc acagtccccg agaagttggg 200gggaggggtc ggcaattgaa
ccggtgccta gagaaggtgg cgcggggtaa 250actgggaaag tgatgtcgtg
tactggctcc gcctttttcc cgagggtggg 300ggagaaccgt atataagtgc
agtagtcgcc gtgaacgttc tttttcgcaa 350cgggtttgcc gccagaacac
aggtaagtgc cgtgtgtggt tcccgcgggc 400ctggcctctt tacgggttat
ggcccttgcg tgccttgaat tacttccacg 450cccctggctg cagtacgtga
ttcttgatcc cgagcttcgg gttggaagtg 500ggtgggagag ttcgaggcct
tgcgcttaag gagccccttc gcctcgtgct 550tgagttgagg cctggcctgg
gcgctggggc cgccgcgtgc gaatctggtg 600gcaccttcgc gcctgtctcg
ctgctttcga taagtctcta gccatttaaa 650atttttgatg acctgctgcg
acgctttttt tctggcaaga tagtcttgta 700aatgcgggcc aagatctgca
cactggtatt tcggtttttg gggccgcggg 750cggcgacggg gcccgtgcgt
cccagcgcac atgttcggcg aggcggggcc 800tgcgagcgcg gccaccgaga
atcggacggg ggtagtctca agctggccgg 850cctgctctgg tgcctggcct
cgcgccgccg tgtatcgccc cgccctgggc 900ggcaaggctg gcccggtcgg
caccagttgc gtgagcggaa agatggccgc 950ttcccggccc tgctgcaggg
agctcaaaat ggaggacgcg gcgctcggga 1000gagcgggcgg gtgagtcacc
cacacaaagg aaaagggcct ttccgtcctc 1050agccgtcgct tcatgtgact
ccacggagta ccgggcgccg tccaggcacc 1100tcgattagtt ctcgagcttt
tggagtacgt cgtctttagg ttggggggag 1150gggttttatg cgatggagtt
tccccacact gagtgggtgg agactgaagt 1200taggccagct tggcacttga
tgtaattctc cttggaattt gccctttttg 1250agtttggatc ttggttcatt
ctcaagcctc agacagtggt tcaaagtttt 1300cttccatttc aggtgtacgc
gtctcgggaa gctttagttt aaacgcctgg 1350atcctctgcg cctgggccca
gctctgtccc acaccgcggt cacatggcac 1400cacctctctt gcagcc tcc acc aag
ggc cca tcg gtc ttc ccc ctg 1446 Ser Thr Lys Gly Pro Ser Val Phe
Pro Leu 1 5 10gca ccc tcc tcc aag agc acc tct ggg ggc aca gcg gcc
ctg ggc 1491Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
Gly 15 20 25tgc ctg gtc aag gac tac ttc ccc gaa ccg gtg acg gtg tcg
tgg 1536Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
30 35 40aac tca ggc gcc ctg acc agc ggc gtg cac acc ttc ccg gct gtc
1581Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 45
50 55cta cag tcc tca gga ctc tac tcc ctc agc agc gtg gtg acc gtg
1626Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Ser Val 60
65 70ccc tcc agc agc ttg ggc acc cag acc tac atc tgc aac gtg aat
1671Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn 75
80 85cac aag ccc agc aac acc aag gtg gac aag aaa gtt gag ccc aaa
1716His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys 90
95 100tct tgt gac aaa act cac aca tgc cca ccg tgc cca gca cct gaa
1761Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu 105
110 115ctc ctg ggg gga ccg tca gtc ttc ctc ttc ccc cca aaa ccc aag
1806Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 120
125 130gac acc ctc atg atc tcc cgg acc cct gag gtc aca tgc gtg gtg
1851Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val 135
140 145gtg gac gtg agc cac gaa gac cct gag gtc aag ttc aac tgg tac
1896Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 150
155 160gtg gac ggc gtg gag gtg cat aac gcc aag aca aag ccg cgg gag
1941Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 165
170 175gag cag tac aac agc acg tac cgg gtg gtc agc gtc ctc acc gtc
1986Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val 180
185 190ctg cac cag gac tgg ctg aat ggc aag gag tac aag tgc aag gtc
2031Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val 195
200 205tcc aac aaa gcc ctc cca gcc ccc atc gag aaa acc atc tcc aaa
2076Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys 210
215 220gcc aaa ggg cag ccc cga gaa cca cag gtg tac acc ctg ccc cca
2121Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro 225
230 235tcc cgg gag gag atg acc aag aac cag gtc agc ctg acc tgc ctg
2166Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu 240
245 250gtc aaa ggc ttc tat ccc agc gac atc gcc gtg gag tgg gag agc
2211Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser 255
260 265aat ggg cag ccg gag aac aac tac aag acc acg cct ccc gtg ctg
2256Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu 270
275 280gac tcc gac ggc tcc ttc ttc ctc tac agc aag ctc acc gtg gac
2301Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 285
290 295aag agc agg tgg cag cag ggg aac gtc ttc tca tgc tcc gtg atg
2346Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met 300
305 310cat gag gct ctg cac aac cac tac acg cag aag agc ctc tcc ctg
2391His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 315
320 325tct ccg ggt aaa tga gctagaaact aactaagcta gcaacggttt 2436Ser
Pro Gly Lysccctctagcg ggatcaattc cgcccccccc ccctaacgtt actggccgaa
2486gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc
2536atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt
2586cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag
2636gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga
2686caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg
2736cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa
2786aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga
2836gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca
2886gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct
2936ttacgtgtgt ttagtcgagg ttaaaaaacg tctaggcccc ccgaaccacg
2986gggacgtggt tttcctttga aaaacacgat aataccatgg ttcgaccatt
3036gaactgcatc gtcgccgtgt cccaaaatat ggggattggc aagaacggag
3086acctaccctg gcctccgctc aggaacgagt tcaagtactt ccaaagaatg
3136accacaacct cttcagtgga aggtaaacag aatctggtga ttatgggtag
3186gaaaacctgg ttctccattc ctgagaagaa tcgaccttta aaggacagaa
3236ttaatggttc gatatagttc tcagtagaga actcaaagaa ccaccacgag
3286gagctcattt tcttgccaaa agtttggatg atgccttaag acttattgaa
3336caaccggaat tggcaagtaa agtagacatg gtttggatag tcggaggcag
3386ttctgtttac caggaagcca tgaatcaacc aggccacctc agactctttg
3436tgacaaggat catgcaggaa tttgaaagtg acacgttttt cccagaaatt
3486gatttgggga aatataaact tctcccagaa tacccaggcg tcctctctga
3536ggtccaggag gaaaaaggca tcaagtataa gtttgaagtc tacgagaaga
3586aagactaaca ggaagatgct ttcaagttct ctgctcccct cctaaagcta
3636tgcattttta taagaccatg ggacttttgc tggtcgatcg acctggcgta
3686atagcgaaga
ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg 3736aatggcgaat
gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt 3786ggtggttacg
cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg 3836ctcctttcgc
tttcttccct tcctttctcg ccacgttcgc cggctttccc 3886cgtcaagctc
taaatcgggg gctcccttta gggttccgat ttagtgcttt 3936acggcacctc
gaccccaaaa aacttgatta gggtgatggt tcacgtagtg 3986ggccatcgcc
ctgatagacg gtttttcgcc tttgacgttg gagtccacgt 4036tctttaatag
tggactcttg ttccaaactg gaacaacact caaccctatc 4086tcggtctatt
tataagggat tttgccgatt tcggcctatt ggttaaaaaa 4136tgagctgatt
taacaaaatt taacgcgaat tttaacaaaa tattaacgct 4186tacaatttag
gtggcacttt tcggggaaat gtgcgcggaa cccctatatt 4236tgtttatttt
tctaaataca ttcaaatatg tatccgctca tgagacaata 4286accctgataa
atgcttcaat aatattgaaa aaggaagagt atgagtattc 4336aacatttccg
tgtcgccctt attccctttt ttgcggcatt ttgccttact 4386gtttttgctc
acccagaaac gctggtgaaa gtaaaagatg ctgaagatca 4436gttgggtgca
cgagtgggtt acatcgaact ggatctcaac agcggtaaga 4486tccttgagag
ttttcgcccc gaagaacgtt ttccaatgat gagcactttt 4536aaagttctgc
tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga 4586gcaactcggt
cgccgcatac actattctca gaatgacttg gttgagtact 4636caccagtcac
agaaaagcat attacggatg gcatgacagt aagagaatta 4686tgcagtgctg
ccataaccat gagtgataac actgcggcca acttacttct 4736gacaacgatc
ggaggaccga aggagctaac cgcttttttg cacaacatgg 4786gggatcatgt
aactcgcctt gatcgttggg aaccggagct gaatgaagcc 4836ataccaaacg
acgagcgtga caccacgatg cctgtagcaa tggcaacaac 4886gttgcgcaaa
ctattaactg gcgaactact tactctagct tcccggcaac 4936aattaataga
ctggatggag gcggataaag ttgcaggacc acttctgcgc 4986tcggcccttc
cggctggctg gtttattgct gataaatctg gagccggtga 5036gcgtgggtct
cgcggtatca ttgcagcact ggggccagat ggtaagccct 5086cccgtatcgt
agttatctac acgacgggga gtcaggcaac tatggatgaa 5136cgaaatagac
agatcgctga gataggtgcc tcactgatta agcattggta 5186actgtcagac
caagtttact catatatact ttagattgat ttaaaacttc 5236atttttaatt
taaaaggatc taggtgaaga tcctttttga taatctcatg 5286accaaaatcc
cttaacgtga gttttcgttc cactgagcgt cagaccccgt 5336agaaaagatc
aaaggatgtt cttgagatcc tttttttctg cacgtaatct 5386gctgcttgca
aacaaaaaac caccgctacc agcggtggtt tgtttgccgg 5436atcaagagct
accaactctt tttccgaagg taactggctt cagcagagcg 5486cagataccaa
atactgtcct tctagtgtag ccgtagttag gccaccactt 5536caagaactct
gtagcaccgc ctacatacct cgctctgcta atcctgttac 5586cagtggctgc
tgccagtggc gataagtcgt gtcttaccgg gttggactca 5636agacgatagt
taccggataa ggcgcagcgg tcgggctgaa cggggggttc 5686gtgcacacag
cccagcttgg agcgaacgac ctacaccgaa ctgagatacc 5736tacagcgtga
gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg 5786gacaggtatc
cggtaagcgg cagggtcgga acaggagagc gcacgaggga 5836gcttccaggg
ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc 5886acctctgact
tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc 5936ctatggaaaa
acgccagcaa cgcggccttt ttacggttcc tggccttttg 5986ctggcctttt
gctcacatgt tctttcctgc gttatcccct gattctgtgg 6036ataaccgtat
taccgccttt gagtgagctg ataccgctcg ccgcagccga 6086acgaccgagc
gcagcgagtc agtgagcgag gaagcggaag agcgcccaat 6136acgcaaaccg
cctctccccg cgcgttggcc gattcattaa tgcaggtatc 6186acgaggccct
ttcgtcttca c 620750444DNAMus musculusmurine G250 heavy chain
variable region 50gtttaaacgc cgccaccatg aacttcgggc tcagattgat
tttccttgtc 50ctggttttaa aaggtgtcct gtgtgacgtg aagctcgtgg agtctggggc
100agccttagtg aagcttggag ggtccctgaa actctcctgt gcagcctctg
150gattcacttt cagtaactat tacatgtctt gggttcgcca gactccagag
200aagaggctgg agttggtcgc agccattaat agtgatggtg gtatcaccta
250ctatctagac actgtgaagg gccgattcac catttcaaga gacaatgcca
300agaacaccct gtacctgcaa atgagcagtc tgaagtctga ggacacagcc
350ttgttttact gtgcaagaca ccgctcaggc tacttttcta tggactactg
400gggtcaagga acctcagtca ccgtctcctc aggtgagtgg atcc 44451140PRTMus
musculusamino acid sequence for murine G250 heavy chain variable
region 51Met Asn Phe Gly Leu Arg Leu Ile Phe Leu Val Leu Val Leu
Lys Gly1 5 10 15Val Leu Cys Asp Val Lys Leu Val Glu Ser Gly Ala Ala
Leu Val Lys 20 25 30Leu Gly Gly Ser Leu Lys Leu Ser Cys Ala Ala Ser
Gly Phe Thr Phe 35 40 45Ser Asn Tyr Tyr Met Ser Trp Val Arg Gln Thr
Pro Glu Lys Arg Leu 50 55 60Glu Leu Val Ala Ala Ile Asn Ser Asp Gly
Gly Ile Thr Tyr Tyr Leu65 70 75 80Asp Thr Val Lys Gly Arg Phe Thr
Ile Ser Arg Asp Asn Ala Lys Asn 85 90 95Thr Leu Tyr Leu Gln Met Ser
Ser Leu Lys Ser Glu Asp Thr Ala Leu 100 105 110Phe Tyr Cys Ala Arg
His Arg Ser Gly Tyr Phe Ser Met Asp Tyr Trp 115 120 125Gly Gln Gly
Thr Ser Val Thr Val Ser Ser Gly Glu 130 135 14052423DNAMus
musculusmurine G250 light chain variable region 52gtttaaacgc
cgccaccatg ggcttcaaga tggagtttca tactcaggtc 50tttgtattcg tgtttctctg
gttgtctggt gttgatggag acattgtgat 100gacccagtct caaagattca
tgtccacaac agtaggagac agggtcagca 150tcacctgcaa ggccagtcag
aatgtggttt ctgctgttgc ctggtatcaa 200cagaaaccag gacaatctcc
taaactactg atttactcag catccaatcg 250gtacactgga gtccctgatc
gcttcacagg cagtggatct gggacagatt 300tcactctcac cattagcaat
atgcagtctg aagacctggc tgattttttc 350tgtcaacaat atagcaacta
tccgtggacg ttcggtggag gcaccaagct 400ggaaatcaaa cgtgagtgga tcc
42353132PRTMus musculusamino acid sequence for murine G250 light
chain variable region 53Met Gly Phe Lys Met Glu Phe His Thr Gln Val
Phe Val Phe Val Phe1 5 10 15Leu Trp Leu Ser Gly Val Asp Gly Asp Ile
Val Met Thr Gln Ser Gln 20 25 30Arg Phe Met Ser Thr Thr Val Gly Asp
Arg Val Ser Ile Thr Cys Lys 35 40 45Ala Ser Gln Asn Val Val Ser Ala
Val Ala Trp Tyr Gln Gln Lys Pro 50 55 60Gly Gln Ser Pro Lys Leu Leu
Ile Tyr Ser Ala Ser Asn Arg Tyr Thr65 70 75 80Gly Val Pro Asp Arg
Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr 85 90 95Leu Thr Ile Ser
Asn Met Gln Ser Glu Asp Leu Ala Asp Phe Phe Cys 100 105 110Gln Gln
Tyr Ser Asn Tyr Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu 115 120
125Glu Ile Lys Arg 13054482DNAHomo sapiensnucleotide sequence of a
TNF fragment 54ccatggtctc atcttctcga accccgagtg acaagcctgt
agcccatgtt 50gtagcaaacc ctcaagctga ggggcagctc cagtggctga accgccgggc
100caatgccctc ctggccaatg gcgtggagct gagagataac cagctggtgg
150tgccatcaga gggcctgtac ctcatctact cccaggtcct cttcaagggc
200caaggctgcc cctccaccca tgtgctcctc acccacacca tcagccgcat
250cgccgtctcc taccagacca aggtcaacct cctctctgcc atcaagagcc
300cctgccagag ggagacccca gagggggctg aggccaagcc ctggtatgag
350cccatctatc tgggaggggt cttccagctg gagaagggtg accgactcag
400cgctgagatc aatcggcccg actatctcga ctttgccgag tctgggcagg
450tctactttgg gatcattgcc ctgtgatcta ga 48255157PRTHomo sapiensamino
acid sequence of a TNF fragment 55Met Val Ser Ser Ser Arg Thr Pro
Ser Asp Lys Pro Val Ala His Val1 5 10 15Val Ala Asn Pro Gln Ala Glu
Gly Gln Leu Gln Trp Leu Asn Arg Arg 20 25 30Ala Asn Ala Leu Leu Ala
Asn Gly Val Glu Leu Arg Asp Asn Gln Leu 35 40 45Val Val Pro Ser Glu
Gly Leu Tyr Leu Ile Tyr Ser Gln Val Leu Phe 50 55 60Lys Gly Gln Gly
Cys Pro Ser Thr His Val Leu Leu Thr His Thr Ile65 70 75 80Ser Arg
Ile Ala Val Ser Tyr Gln Thr Lys Val Asn Leu Leu Ser Ala 85 90 95Ile
Lys Ser Pro Cys Gln Arg Glu Thr Pro Glu Gly Ala Glu Ala Lys 100 105
110Pro Trp Tyr Glu Pro Ile Tyr Leu Gly Gly Val Phe Gln Leu Glu Lys
115 120 125Gly Asp Arg Leu Ser Ala Glu Ile Asn Arg Pro Asp Tyr Leu
Asp Phe 130 135 140Ala Glu Ser Gly Gln Val Tyr Phe Gly Ile Ile Ala
Leu145 150 155566047DNAArtificial SequenceHeavy chain expression
vector pREN-DHFR-TNF 56ctcgagagcg ggcagtgagc gcaacgcaat taatgtgagt
tagctcactc 50attaggcacc ccaggcttta cactttatgc tcccggctcg tatgttgtgt
100ggagattgtg agcggataac aatttcacac agaattcgtg aggctccggt
150gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
200gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa
250actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg
300ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa
350cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc
400ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccacg
450cccctggctg cagtacgtga ttcttgatcc cgagcttcgg gttggaagtg
500ggtgggagag ttcgaggcct tgcgcttaag gagccccttc gcctcgtgct
550tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc gaatctggtg
600gcaccttcgc gcctgtctcg ctgctttcga taagtctcta gccatttaaa
650atttttgatg acctgctgcg acgctttttt tctggcaaga tagtcttgta
700aatgcgggcc aagatctgca cactggtatt tcggtttttg gggccgcggg
750cggcgacggg gcccgtgcgt cccagcgcac atgttcggcg aggcggggcc
800tgcgagcgcg gccaccgaga atcggacggg ggtagtctca agctggccgg
850cctgctctgg tgcctggcct cgcgccgccg tgtatcgccc cgccctgggc
900ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa agatggccgc
950ttcccggccc tgctgcaggg agctcaaaat ggaggacgcg gcgctcggga
1000gagcgggcgg gtgagtcacc cacacaaagg aaaagggcct ttccgtcctc
1050agccgtcgct tcatgtgact ccacggagta ccgggcgccg tccaggcacc
1100tcgattagtt ctcgagcttt tggagtacgt cgtctttagg ttggggggag
1150gggttttatg cgatggagtt tccccacact gagtgggtgg agactgaagt
1200taggccagct tggcacttga tgtaattctc cttggaattt gccctttttg
1250agtttggatc ttggttcatt ctcaagcctc agacagtggt tcaaagtttt
1300tttcttccat ttcaggtgta cgcgtctcgg gaagctttag tttaaacgcc
1350ggatcctctg cgcctgggcc cagctctgtc ccacaccgcg gtcacatggc
1400accacctctc ttgcagcc tcc acc aag ggc cca tcg gtc ttc ccc ctg
1448 Ser Thr Lys Gly Pro Ser Val Phe Pro Leu 1 5 10gca ccc tcc tcc
aag agc acc tct ggg ggc aca gcg gcc ctg ggc 1493Ala Pro Ser Ser Lys
Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly 15 20 25tgc ctg gtc aag gac
tac ttc ccc gaa ccg gtg acg gtg tcg tgg 1538Cys Leu Val Lys Asp Tyr
Phe Pro Glu Pro Val Thr Val Ser Trp 30 35 40aac tca ggc gcc ctg acc
agc ggc gtg cac acc ttc ccg gct gtc 1583Asn Ser Gly Ala Leu Thr Ser
Gly Val His Thr Phe Pro Ala Val 45 50 55cta cag tcc tca gga ctc tac
tcc ctc agc agc gtg gtg acc gtg 1628Leu Gln Ser Ser Gly Leu Tyr Ser
Leu Ser Ser Val Val Ser Val 60 65 70ccc tcc agc agc ttg ggc acc cag
acc tac atc tgc aac gtg aat 1673Pro Ser Ser Ser Leu Gly Thr Gln Thr
Tyr Ile Cys Asn Val Asn 75 80 85cac aag ccc agc aac acc aag gtg gac
aag aaa gtt gag ccc aaa 1718His Lys Pro Ser Asn Thr Lys Val Asp Lys
Lys Val Glu Pro Lys 90 95 100tct tgt gac aaa act cac aca tgc cca
ccg tgc cca ggt gga ggt 1763Ser Cys Asp Lys Thr His Thr Cys Pro Pro
Cys Pro Gly Gly Gly 105 110 115gga tca cca atg gtc tca tct tct cga
acc ccg agt gac aag cct 1808Gly Ser Pro Met Val Ser Ser Ser Arg Thr
Pro Ser Asp Lys Pro 120 125 130gta gcc cat gtt gta gca aac cct caa
gct gag ggg cag ctc cag 1853Val Ala His Val Val Ala Asn Pro Gln Ala
Glu Gly Gln Leu Gln 135 140 145tgg ctg aac cgc cgg gcc aat gcc ctc
ctg gcc aat ggc gtg gag 1898Trp Leu Asn Arg Arg Ala Asn Ala Leu Leu
Ala Asn Glu Val Glu 150 155 160ctg aga gat aac cag ctg gtg gtg cca
tca gag ggc ctg tac ctc 1943Leu Arg Asp Asn Gln Leu Val Val Pro Ser
Glu Gly Leu Tyr Leu 165 170 175atc tac tcc cag gtc ctc ttc aag ggc
caa ggc tgc ccc tcc acc 1988Ile Tyr Ser Gln Val Leu Phe Lys Gly Gln
Gly Cys Pro Ser Thr 180 185 190cat gtg ctc ctc acc cac acc atc agc
cgc atc gcc gtc tcc tac 2033His Val Leu Leu Thr His Thr Ile Ser Arg
Ile Ala Val Ser Tyr 195 200 205cag acc aag gtc aac ctc ctc tct gcc
atc aag agc ccc tgc cag 2078Gln Thr Lys Val Asn Leu Leu Ser Ala Ile
Lys Ser Pro Cys Gln 210 215 220agg gag acc cca gag ggg gct gag gcc
aag ccc tgg tat gag ccc 2123Arg Glu Thr Pro Glu Gly Ala Glu Ala Lys
Pro Trp Tyr Glu Pro 225 230 235atc tat ctg gga ggg gtc ttc cag ctg
gag aag ggt gac cga ctc 2168Ile Tyr Leu Gly Gly Val Phe Gln Leu Glu
Lys Gly Asp Arg Leu 240 245 250agc gct gag atc aat cgg ccc gac tat
ctc gac ttt gcc gag tct 2213Ser Ala Glu Ile Asn Arg Pro Asp Tyr Leu
Asp Phe Ala Glu Ser 255 260 265ggg cag gtc tac ttt ggg atc att gcc
ctg tga tctagaaact 2256Gly Gln Val Tyr Phe Gly Ile Ile Ala Leu 270
275aactaagcta gcaacggttt ccctctagcg ggatcaattc cgcccccccc
2306ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg
2356tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc
2406cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc
2456tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc
2506ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg
2556cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg
2606tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga
2656gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac
2706aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct
2756ggggcctcgg tgcacatgct ttacgtgtgt ttagtcgagg ttaaaaaacg
2806tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat
2856aataccatgg ttcgaccatt gaactgcatc gtcgccgtgt cccaaaatat
2906ggggattggc aagaacggag acctaccctg gcctccgctc aggaacgagt
2956tcaagtactt ccaaagaatg accacaacct cttcagtgga aggtaaacag
3006aatctggtga ttatgggtag gaaaacctgg ttctccattc ctgagaagaa
3056tcgaccttta aaggacagaa ttaatggttc gatatagttc tcagtagaga
3106actcaaagaa ccaccacgag gagctcattt tcttgccaaa agtttggatg
3156atgccttaag acttattgaa caaccggaat tggcaagtaa agtagacatg
3206gtttggatag tcggaggcag ttctgtttac caggaagcca tgaatcaacc
3256aggccacctc agactctttg tgacaaggat catgcaggaa tttgaaagtg
3306acacgttttt cccagaaatt gatttgggga aatataaact tctcccagaa
3356tacccaggcg tcctctctga ggtccaggag gaaaaaggca tcaagtataa
3406gtttgaagtc tacgagaaga aagactaaca ggaagatgct ttcaagttct
3456ctgctcccct cctaaagcta tgcattttta taagaccatg ggacttttgc
3506tggtcgatcg acctggcgta atagcgaaga ggcccgcacc gatcgccctt
3556cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc
3606gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
3656tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
3706ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
3756gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
3806gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
3856tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
3906gaacaacact caaccctatc tcggtctatt tataagggat tttgccgatt
3956tcggcctatt ggttaaaaaa tgagctgatt taacaaaatt taacgcgaat
4006tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat
4056gtgcgcggaa cccctatatt tgtttatttt tctaaataca ttcaaatatg
4106tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
4156aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
4206ttgcggcatt ttgccttact gtttttgctc acccagaaac gctggtgaaa
4256gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
4306ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
4356ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
4406cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
4456gaatgacttg gttgagtact caccagtcac agaaaagcat attacggatg
4506gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
4556actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
4606cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
4656aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
4706cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
4756tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
4806ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
4856gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
4906ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
4956gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
5006tcactgatta agcattggta actgtcagac caagtttact catatatact
5056ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
5106tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
5156cactgagcgt cagaccccgt agaaaagatc aaaggatgtt cttgagatcc
5206tttttttctg cacgtaatct gctgcttgca aacaaaaaac caccgctacc
5256agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
5306taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
5356ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
5406cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
5456gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
5506tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
5556ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
5606ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
5656acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
5706tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
5756gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
5806ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
5856gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
5906ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
5956gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
6006gattcattaa tgcaggtatc acgaggccct ttcgtcttca c 6047
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