U.S. patent application number 13/844818 was filed with the patent office on 2013-10-03 for antibody-mediated transduction of heat shock proteins into living cells. The applicant listed for this patent is Robert N. Nishimura, Richard H. Weisbart. Invention is credited to Robert N. Nishimura, Richard H. Weisbart.
Application Number | 20130259862 13/844818 |
Document ID | / |
Family ID | 49235341 |
Filed Date | 2013-10-03 |
United States Patent Application | 20130259862 |
Kind Code | A1 |
Nishimura; Robert N. ; et al. | October 3, 2013 |
The invention provides for a fusion protein comprising a 3E10 Fv joined to a Hsp-70, Hsp-27, Hsp-90 or GRP-78 or portion thereof, and optionally, the 3E10 Fv comprising an amino acid sequence AGIH at its amino terminus.
Inventors: | Nishimura; Robert N.; (Sepulveda, CA) ; Weisbart; Richard H.; (Sepulveda, CA) | ||||||||||
Applicant: |
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Family ID: | 49235341 | ||||||||||
Appl. No.: | 13/844818 | ||||||||||
Filed: | March 16, 2013 |
Application Number | Filing Date | Patent Number | ||
---|---|---|---|---|
61618594 | Mar 30, 2012 | |||
Current U.S. Class: | 424/134.1 ; 530/387.3 |
Current CPC Class: | C07K 2317/56 20130101; C07K 2317/77 20130101; C07K 2317/622 20130101; C07K 2319/33 20130101; C07K 16/18 20130101; A61K 38/00 20130101; C07K 14/47 20130101 |
Class at Publication: | 424/134.1 ; 530/387.3 |
International Class: | A61K 47/48 20060101 A61K047/48 |
Sequence CWU 1
1
3611764DNAArtificial Sequence3E10 Fv-Hsp27 nucleotide sequence
encoding a chimeric protein comprising a single chain Fv antibody
fragment derived from Mus musculus 3E10 hybridoma and Hsp27 from
Homo sapiens joined by mouse CH1 linker and swivel sequence 1atg
aga ttt cct tca ctt ttt act gct gtt tta ttc gca gca tcc tcc 48Met
Arg Phe Pro Ser Leu Phe Thr Ala Val Leu Phe Ala Ala Ser Ser 1 5 10
15 gca tta gct gct cca gtc aac act aca aca gaa gat gaa acg gca caa
96Ala Leu Ala Ala Pro Val Asn Thr Thr Thr Glu Asp Glu Thr Ala Gln
20 25 30 att ccg gct gaa gct gtc atc ggt tac tca gat tta gaa ggg
gat ttc 144Ile Pro Ala Glu Ala Val Ile Gly Tyr Ser Asp Leu Glu Gly
Asp Phe 35 40 45 gat gtt gct gtt ttg cca ttt tcc aac agc aca aat
aac ggg tta ttg 192Asp Val Ala Val Leu Pro Phe Ser Asn Ser Thr Asn
Asn Gly Leu Leu 50 55 60 ttt ata aat act act att gcc agc att gct
gct aaa gaa gaa ggg gta 240Phe Ile Asn Thr Thr Ile Ala Ser Ile Ala
Ala Lys Glu Glu Gly Val 65 70 75 80 tct ctc gag aaa aga gag gct gaa
gct gaa ttc cat cac cat cac cat 288Ser Leu Glu Lys Arg Glu Ala Glu
Ala Glu Phe His His His His His 85 90 95 cac gca ggg att cac gac
att gtc ctg aca cag tct cct gct tcc tta 336His Ala Gly Ile His Asp
Ile Val Leu Thr Gln Ser Pro Ala Ser Leu 100 105 110 gct gta tct ctg
ggg cag agg gcc acc atc tcc tgc agg gcc agc aaa 384Ala Val Ser Leu
Gly Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Lys 115 120 125 agt gtc
agt aca tct agc tat agt tac atg cac tgg tac caa cag aaa 432Ser Val
Ser Thr Ser Ser Tyr Ser Tyr Met His Trp Tyr Gln Gln Lys 130 135 140
cca gga cag cca ccc aaa ctc ctc atc aag tat gca tcc tac cta gaa
480Pro Gly Gln Pro Pro Lys Leu Leu Ile Lys Tyr Ala Ser Tyr Leu Glu
145 150 155 160 tct ggg gtt cct gcc agg ttc agt ggc agt ggg tct ggg
aca gac ttc 528Ser Gly Val Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly
Thr Asp Phe 165 170 175 acc ctc aac atc cat cct gtg gag gag gag gat
gct gca aca tat tac 576Thr Leu Asn Ile His Pro Val Glu Glu Glu Asp
Ala Ala Thr Tyr Tyr 180 185 190 tgt cag cac agt agg gag ttt ccg tgg
acg ttc ggt gga ggc acc aag 624Cys Gln His Ser Arg Glu Phe Pro Trp
Thr Phe Gly Gly Gly Thr Lys 195 200 205 ctg gaa atc aaa cgg gct gat
gct gca ccc ggg ggt ggc ggt tct ggc 672Leu Glu Ile Lys Arg Ala Asp
Ala Ala Pro Gly Gly Gly Gly Ser Gly 210 215 220 ggt ggc ggt tct gga
ggc ggt ggc tct gag gtg cag ctg gtg gag tct 720Gly Gly Gly Ser Gly
Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser 225 230 235 240 ggg gga
ggc tta gtg aag cct gga ggg tcc cgg aaa ctc tcc tgt gca 768Gly Gly
Gly Leu Val Lys Pro Gly Gly Ser Arg Lys Leu Ser Cys Ala 245 250 255
gcc tct gga ttc act ttc agt aac tat gga atg cac tgg gtc cgt cag
816Ala Ser Gly Phe Thr Phe Ser Asn Tyr Gly Met His Trp Val Arg Gln
260 265 270 gct cca gag aag ggg ctg gag tgg gtt gca tac att agt agt
ggc agt 864Ala Pro Glu Lys Gly Leu Glu Trp Val Ala Tyr Ile Ser Ser
Gly Ser 275 280 285 agt acc atc tac tat gca gac aca gtg aag ggc cga
ttc acc atc tcc 912Ser Thr Ile Tyr Tyr Ala Asp Thr Val Lys Gly Arg
Phe Thr Ile Ser 290 295 300 aga gac aat gcc aag aac acc ctg ttc ctg
caa atg acc agt cta agg 960Arg Asp Asn Ala Lys Asn Thr Leu Phe Leu
Gln Met Thr Ser Leu Arg 305 310 315 320 tct gag gac aca gcc atg cgg
ggg tta cta ctt gac tac tgg ggc caa 1008Ser Glu Asp Thr Ala Met Arg
Gly Leu Leu Leu Asp Tyr Trp Gly Gln 325 330 335 ggc acc act ctc aca
gtc tcc tca cta gaa caa aaa ctc atc tca gaa 1056Gly Thr Thr Leu Thr
Val Ser Ser Leu Glu Gln Lys Leu Ile Ser Glu 340 345 350 gag gat ctg
aat agc gcc gtc gac gcc aaa aca aca gcc cca tcg gtc 1104Glu Asp Leu
Asn Ser Ala Val Asp Ala Lys Thr Thr Ala Pro Ser Val 355 360 365 tat
cca ctg gcc cct gta ctg gag tct tcc gga tcc atg acc gag cgc 1152Tyr
Pro Leu Ala Pro Val Leu Glu Ser Ser Gly Ser Met Thr Glu Arg 370 375
380 cgc gtc ccc ttc tcg ctc ctg cgg ggc ccc agc tgg gac ccc ttc cgc
1200Arg Val Pro Phe Ser Leu Leu Arg Gly Pro Ser Trp Asp Pro Phe Arg
385 390 395 400 gac tgg tac ccg cat agc cgc ctc ttc gac cag gcc ttc
ggg ctg ccc 1248Asp Trp Tyr Pro His Ser Arg Leu Phe Asp Gln Ala Phe
Gly Leu Pro 405 410 415 cgg ctg ccg gag gag tgg tcg cag tgg tta ggc
ggc agc agc tgg cca 1296Arg Leu Pro Glu Glu Trp Ser Gln Trp Leu Gly
Gly Ser Ser Trp Pro 420 425 430 ggc tac gtg cgc ccc ctg ccc ccc gcc
gcc atc gag agc ccc gca gtg 1344Gly Tyr Val Arg Pro Leu Pro Pro Ala
Ala Ile Glu Ser Pro Ala Val 435 440 445 gcc gcg ccc gcc tac agc cgc
gcg ctc agc cgg caa ctc agc agc ggg 1392Ala Ala Pro Ala Tyr Ser Arg
Ala Leu Ser Arg Gln Leu Ser Ser Gly 450 455 460 gtc tcg gag atc cgg
cac act gcg gac cgc tgg cgc gtg tcc ctg gat 1440Val Ser Glu Ile Arg
His Thr Ala Asp Arg Trp Arg Val Ser Leu Asp 465 470 475 480 gtc aac
cac ttc gcc ccg gac gag ctg acg gtc aag acc aag gat ggc 1488Val Asn
His Phe Ala Pro Asp Glu Leu Thr Val Lys Thr Lys Asp Gly 485 490 495
gtg gtg gag atc acc ggc aag cac gag gag cgg cag gac gag cat ggc
1536Val Val Glu Ile Thr Gly Lys His Glu Glu Arg Gln Asp Glu His Gly
500 505 510 tac atc tcc cgg tgc ttc acg cgg aaa tac acg ctg ccc ccc
ggt gtg 1584Tyr Ile Ser Arg Cys Phe Thr Arg Lys Tyr Thr Leu Pro Pro
Gly Val 515 520 525 gac ccc acc caa gtt tcc tcc tcc ctg tcc cct gag
ggc aca ctg acc 1632Asp Pro Thr Gln Val Ser Ser Ser Leu Ser Pro Glu
Gly Thr Leu Thr 530 535 540 gtg gag gcc ccc atg ccc aag cta gcc acg
cag tcc aac gag atc acc 1680Val Glu Ala Pro Met Pro Lys Leu Ala Thr
Gln Ser Asn Glu Ile Thr 545 550 555 560 atc cca gtc acc ttc gag tcg
cgg gcc cag ctt ggg ggc cca gaa gct 1728Ile Pro Val Thr Phe Glu Ser
Arg Ala Gln Leu Gly Gly Pro Glu Ala 565 570 575 gca aaa tcc gat gag
act gcc gcc aag taa tctaga 1764Ala Lys Ser Asp Glu Thr Ala Ala Lys
580 585 2585PRTArtificial SequenceSynthetic Construct 2Met Arg Phe
Pro Ser Leu Phe Thr Ala Val Leu Phe Ala Ala Ser Ser 1 5 10 15 Ala
Leu Ala Ala Pro Val Asn Thr Thr Thr Glu Asp Glu Thr Ala Gln 20 25
30 Ile Pro Ala Glu Ala Val Ile Gly Tyr Ser Asp Leu Glu Gly Asp Phe
35 40 45 Asp Val Ala Val Leu Pro Phe Ser Asn Ser Thr Asn Asn Gly
Leu Leu 50 55 60 Phe Ile Asn Thr Thr Ile Ala Ser Ile Ala Ala Lys
Glu Glu Gly Val 65 70 75 80 Ser Leu Glu Lys Arg Glu Ala Glu Ala Glu
Phe His His His His His 85 90 95 His Ala Gly Ile His Asp Ile Val
Leu Thr Gln Ser Pro Ala Ser Leu 100 105 110 Ala Val Ser Leu Gly Gln
Arg Ala Thr Ile Ser Cys Arg Ala Ser Lys 115 120 125 Ser Val Ser Thr
Ser Ser Tyr Ser Tyr Met His Trp Tyr Gln Gln Lys 130 135 140 Pro Gly
Gln Pro Pro Lys Leu Leu Ile Lys Tyr Ala Ser Tyr Leu Glu 145 150 155
160 Ser Gly Val Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe
165 170 175 Thr Leu Asn Ile His Pro Val Glu Glu Glu Asp Ala Ala Thr
Tyr Tyr 180 185 190 Cys Gln His Ser Arg Glu Phe Pro Trp Thr Phe Gly
Gly Gly Thr Lys 195 200 205 Leu Glu Ile Lys Arg Ala Asp Ala Ala Pro
Gly Gly Gly Gly Ser Gly 210 215 220 Gly Gly Gly Ser Gly Gly Gly Gly
Ser Glu Val Gln Leu Val Glu Ser 225 230 235 240 Gly Gly Gly Leu Val
Lys Pro Gly Gly Ser Arg Lys Leu Ser Cys Ala 245 250 255 Ala Ser Gly
Phe Thr Phe Ser Asn Tyr Gly Met His Trp Val Arg Gln 260 265 270 Ala
Pro Glu Lys Gly Leu Glu Trp Val Ala Tyr Ile Ser Ser Gly Ser 275 280
285 Ser Thr Ile Tyr Tyr Ala Asp Thr Val Lys Gly Arg Phe Thr Ile Ser
290 295 300 Arg Asp Asn Ala Lys Asn Thr Leu Phe Leu Gln Met Thr Ser
Leu Arg 305 310 315 320 Ser Glu Asp Thr Ala Met Arg Gly Leu Leu Leu
Asp Tyr Trp Gly Gln 325 330 335 Gly Thr Thr Leu Thr Val Ser Ser Leu
Glu Gln Lys Leu Ile Ser Glu 340 345 350 Glu Asp Leu Asn Ser Ala Val
Asp Ala Lys Thr Thr Ala Pro Ser Val 355 360 365 Tyr Pro Leu Ala Pro
Val Leu Glu Ser Ser Gly Ser Met Thr Glu Arg 370 375 380 Arg Val Pro
Phe Ser Leu Leu Arg Gly Pro Ser Trp Asp Pro Phe Arg 385 390 395 400
Asp Trp Tyr Pro His Ser Arg Leu Phe Asp Gln Ala Phe Gly Leu Pro 405
410 415 Arg Leu Pro Glu Glu Trp Ser Gln Trp Leu Gly Gly Ser Ser Trp
Pro 420 425 430 Gly Tyr Val Arg Pro Leu Pro Pro Ala Ala Ile Glu Ser
Pro Ala Val 435 440 445 Ala Ala Pro Ala Tyr Ser Arg Ala Leu Ser Arg
Gln Leu Ser Ser Gly 450 455 460 Val Ser Glu Ile Arg His Thr Ala Asp
Arg Trp Arg Val Ser Leu Asp 465 470 475 480 Val Asn His Phe Ala Pro
Asp Glu Leu Thr Val Lys Thr Lys Asp Gly 485 490 495 Val Val Glu Ile
Thr Gly Lys His Glu Glu Arg Gln Asp Glu His Gly 500 505 510 Tyr Ile
Ser Arg Cys Phe Thr Arg Lys Tyr Thr Leu Pro Pro Gly Val 515 520 525
Asp Pro Thr Gln Val Ser Ser Ser Leu Ser Pro Glu Gly Thr Leu Thr 530
535 540 Val Glu Ala Pro Met Pro Lys Leu Ala Thr Gln Ser Asn Glu Ile
Thr 545 550 555 560 Ile Pro Val Thr Phe Glu Ser Arg Ala Gln Leu Gly
Gly Pro Glu Ala 565 570 575 Ala Lys Ser Asp Glu Thr Ala Ala Lys 580
585 3585PRTArtificial Sequence3E10 Fv-Hsp27 amino acid sequence for
a chimeric protein comprising a single chain Fv antibody fragment
derived from Mus musculus 3E10 hybridoma and Hsp27 from Homo
sapiens joined by Mus musculus CH1 linker and swivel sequence 3Met
Arg Phe Pro Ser Leu Phe Thr Ala Val Leu Phe Ala Ala Ser Ser 1 5 10
15 Ala Leu Ala Ala Pro Val Asn Thr Thr Thr Glu Asp Glu Thr Ala Gln
20 25 30 Ile Pro Ala Glu Ala Val Ile Gly Tyr Ser Asp Leu Glu Gly
Asp Phe 35 40 45 Asp Val Ala Val Leu Pro Phe Ser Asn Ser Thr Asn
Asn Gly Leu Leu 50 55 60 Phe Ile Asn Thr Thr Ile Ala Ser Ile Ala
Ala Lys Glu Glu Gly Val 65 70 75 80 Ser Leu Glu Lys Arg Glu Ala Glu
Ala Glu Phe His His His His His 85 90 95 His Ala Gly Ile His Asp
Ile Val Leu Thr Gln Ser Pro Ala Ser Leu 100 105 110 Ala Val Ser Leu
Gly Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Lys 115 120 125 Ser Val
Ser Thr Ser Ser Tyr Ser Tyr Met His Trp Tyr Gln Gln Lys 130 135 140
Pro Gly Gln Pro Pro Lys Leu Leu Ile Lys Tyr Ala Ser Tyr Leu Glu 145
150 155 160 Ser Gly Val Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr
Asp Phe 165 170 175 Thr Leu Asn Ile His Pro Val Glu Glu Glu Asp Ala
Ala Thr Tyr Tyr 180 185 190 Cys Gln His Ser Arg Glu Phe Pro Trp Thr
Phe Gly Gly Gly Thr Lys 195 200 205 Leu Glu Ile Lys Arg Ala Asp Ala
Ala Pro Gly Gly Gly Gly Ser Gly 210 215 220 Gly Gly Gly Ser Gly Gly
Gly Gly Ser Glu Val Gln Leu Val Glu Ser 225 230 235 240 Gly Gly Gly
Leu Val Lys Pro Gly Gly Ser Arg Lys Leu Ser Cys Ala 245 250 255 Ala
Ser Gly Phe Thr Phe Ser Asn Tyr Gly Met His Trp Val Arg Gln 260 265
270 Ala Pro Glu Lys Gly Leu Glu Trp Val Ala Tyr Ile Ser Ser Gly Ser
275 280 285 Ser Thr Ile Tyr Tyr Ala Asp Thr Val Lys Gly Arg Phe Thr
Ile Ser 290 295 300 Arg Asp Asn Ala Lys Asn Thr Leu Phe Leu Gln Met
Thr Ser Leu Arg 305 310 315 320 Ser Glu Asp Thr Ala Met Arg Gly Leu
Leu Leu Asp Tyr Trp Gly Gln 325 330 335 Gly Thr Thr Leu Thr Val Ser
Ser Leu Glu Gln Lys Leu Ile Ser Glu 340 345 350 Glu Asp Leu Asn Ser
Ala Val Asp Ala Lys Thr Thr Ala Pro Ser Val 355 360 365 Tyr Pro Leu
Ala Pro Val Leu Glu Ser Ser Gly Ser Met Thr Glu Arg 370 375 380 Arg
Val Pro Phe Ser Leu Leu Arg Gly Pro Ser Trp Asp Pro Phe Arg 385 390
395 400 Asp Trp Tyr Pro His Ser Arg Leu Phe Asp Gln Ala Phe Gly Leu
Pro 405 410 415 Arg Leu Pro Glu Glu Trp Ser Gln Trp Leu Gly Gly Ser
Ser Trp Pro 420 425 430 Gly Tyr Val Arg Pro Leu Pro Pro Ala Ala Ile
Glu Ser Pro Ala Val 435 440 445 Ala Ala Pro Ala Tyr Ser Arg Ala Leu
Ser Arg Gln Leu Ser Ser Gly 450 455 460 Val Ser Glu Ile Arg His Thr
Ala Asp Arg Trp Arg Val Ser Leu Asp 465 470 475 480 Val Asn His Phe
Ala Pro Asp Glu Leu Thr Val Lys Thr Lys Asp Gly 485 490 495 Val Val
Glu Ile Thr Gly Lys His Glu Glu Arg Gln Asp Glu His Gly 500 505 510
Tyr Ile Ser Arg Cys Phe Thr Arg Lys Tyr Thr Leu Pro Pro Gly Val 515
520 525 Asp Pro Thr Gln Val Ser Ser Ser Leu Ser Pro Glu Gly Thr Leu
Thr 530 535 540 Val Glu Ala Pro Met Pro Lys Leu Ala Thr Gln Ser Asn
Glu Ile Thr 545 550 555 560 Ile Pro Val Thr Phe Glu Ser Arg Ala Gln
Leu Gly Gly Pro Glu Ala 565 570 575 Ala Lys Ser Asp Glu Thr Ala Ala
Lys 580 585 43072DNAArtificial Sequence3E10 Fv-Hsp70 nucleotide
sequence encoding a chimeric protein comprising a single chain Fv
antibody fragment derived from Mus musculus 3E10 hybridoma and
Hsp70 from Homo sapiens joined by mouse CH1 linker and swivel
sequence 4atg aga ttt cct tca ctt ttt act gct gtt tta ttc gca gca
tcc tcc
48Met Arg Phe Pro Ser Leu Phe Thr Ala Val Leu Phe Ala Ala Ser Ser 1
5 10 15 gca tta gct gct cca gtc aac act aca aca gaa gat gaa acg gca
caa 96Ala Leu Ala Ala Pro Val Asn Thr Thr Thr Glu Asp Glu Thr Ala
Gln 20 25 30 att ccg gct gaa gct gtc atc ggt tac tca gat tta gaa
ggg gat ttc 144Ile Pro Ala Glu Ala Val Ile Gly Tyr Ser Asp Leu Glu
Gly Asp Phe 35 40 45 gat gtt gct gtt ttg cca ttt tcc aac agc aca
aat aac ggg tta ttg 192Asp Val Ala Val Leu Pro Phe Ser Asn Ser Thr
Asn Asn Gly Leu Leu 50 55 60 ttt ata aat act act att gcc agc att
gct gct aaa gaa gaa ggg gta 240Phe Ile Asn Thr Thr Ile Ala Ser Ile
Ala Ala Lys Glu Glu Gly Val 65 70 75 80 tct ctc gag aaa aga gag gct
gaa gct gaa ttc cat cac cat cac cat 288Ser Leu Glu Lys Arg Glu Ala
Glu Ala Glu Phe His His His His His 85 90 95 cac gca ggg att cac
gac att gtc ctg aca cag tct cct gct tcc tta 336His Ala Gly Ile His
Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu 100 105 110 gct gta tct
ctg ggg cag agg gcc acc atc tcc tgc agg gcc agc aaa 384Ala Val Ser
Leu Gly Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Lys 115 120 125 agt
gtc agt aca tct agc tat agt tac atg cac tgg tac caa cag aaa 432Ser
Val Ser Thr Ser Ser Tyr Ser Tyr Met His Trp Tyr Gln Gln Lys 130 135
140 cca gga cag cca ccc aaa ctc ctc atc aag tat gca tcc tac cta gaa
480Pro Gly Gln Pro Pro Lys Leu Leu Ile Lys Tyr Ala Ser Tyr Leu Glu
145 150 155 160 tct ggg gtt cct gcc agg ttc agt ggc agt ggg tct ggg
aca gac ttc 528Ser Gly Val Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly
Thr Asp Phe 165 170 175 acc ctc aac atc cat cct gtg gag gag gag gat
gct gca aca tat tac 576Thr Leu Asn Ile His Pro Val Glu Glu Glu Asp
Ala Ala Thr Tyr Tyr 180 185 190 tgt cag cac agt agg gag ttt ccg tgg
acg ttc ggt gga ggc acc aag 624Cys Gln His Ser Arg Glu Phe Pro Trp
Thr Phe Gly Gly Gly Thr Lys 195 200 205 ctg gaa atc aaa cgg gct gat
gct gca ccc ggg ggt ggc ggt tct ggc 672Leu Glu Ile Lys Arg Ala Asp
Ala Ala Pro Gly Gly Gly Gly Ser Gly 210 215 220 ggt ggc ggt tct gga
ggc ggt ggc tct gag gtg cag ctg gtg gag tct 720Gly Gly Gly Ser Gly
Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser 225 230 235 240 ggg gga
ggc tta gtg aag cct gga ggg tcc cgg aaa ctc tcc tgt gca 768Gly Gly
Gly Leu Val Lys Pro Gly Gly Ser Arg Lys Leu Ser Cys Ala 245 250 255
gcc tct gga ttc act ttc agt aac tat gga atg cac tgg gtc cgt cag
816Ala Ser Gly Phe Thr Phe Ser Asn Tyr Gly Met His Trp Val Arg Gln
260 265 270 gct cca gag aag ggg ctg gag tgg gtt gca tac att agt agt
ggc agt 864Ala Pro Glu Lys Gly Leu Glu Trp Val Ala Tyr Ile Ser Ser
Gly Ser 275 280 285 agt acc atc tac tat gca gac aca gtg aag ggc cga
ttc acc atc tcc 912Ser Thr Ile Tyr Tyr Ala Asp Thr Val Lys Gly Arg
Phe Thr Ile Ser 290 295 300 aga gac aat gcc aag aac acc ctg ttc ctg
caa atg acc agt cta agg 960Arg Asp Asn Ala Lys Asn Thr Leu Phe Leu
Gln Met Thr Ser Leu Arg 305 310 315 320 tct gag gac aca gcc atg cgg
ggg tta cta ctt gac tac tgg ggc caa 1008Ser Glu Asp Thr Ala Met Arg
Gly Leu Leu Leu Asp Tyr Trp Gly Gln 325 330 335 ggc acc act ctc aca
gtc tcc tca cta gaa caa aaa ctc atc tca gaa 1056Gly Thr Thr Leu Thr
Val Ser Ser Leu Glu Gln Lys Leu Ile Ser Glu 340 345 350 gag gat ctg
aat agc gcc gtc gac gcc aaa aca aca gcc cca tcg gtc 1104Glu Asp Leu
Asn Ser Ala Val Asp Ala Lys Thr Thr Ala Pro Ser Val 355 360 365 tat
cca ctg gcc cct gta ctg gag tct tcc gga tcc atg gcc aaa gcc 1152Tyr
Pro Leu Ala Pro Val Leu Glu Ser Ser Gly Ser Met Ala Lys Ala 370 375
380 gcg gcg atc ggc atc gac ctg ggc acc acc tac tcc tgc gtg ggg gtg
1200Ala Ala Ile Gly Ile Asp Leu Gly Thr Thr Tyr Ser Cys Val Gly Val
385 390 395 400 ttc caa cac ggc aag gtg gag atc atc gcc aac gac cag
ggc aac cgc 1248Phe Gln His Gly Lys Val Glu Ile Ile Ala Asn Asp Gln
Gly Asn Arg 405 410 415 acc acc ccc agc tac gtg gcc ttc acg gac acc
gag cgg ctc atc ggg 1296Thr Thr Pro Ser Tyr Val Ala Phe Thr Asp Thr
Glu Arg Leu Ile Gly 420 425 430 gat gcg gcc aag aac cag gtg gcg ctg
aac ccg cag aac acc gtg ttt 1344Asp Ala Ala Lys Asn Gln Val Ala Leu
Asn Pro Gln Asn Thr Val Phe 435 440 445 gac gcg aag cgg ctg atc ggc
cgc aag ttc ggc gac ccg gtg gtg cag 1392Asp Ala Lys Arg Leu Ile Gly
Arg Lys Phe Gly Asp Pro Val Val Gln 450 455 460 tcg gac atg aag cac
tgg cct ttc cag gtg atc aac gac gga gac aag 1440Ser Asp Met Lys His
Trp Pro Phe Gln Val Ile Asn Asp Gly Asp Lys 465 470 475 480 ccc aag
gtg cag gtg agc tac aag ggg gag acc aag gca ttc tac ccc 1488Pro Lys
Val Gln Val Ser Tyr Lys Gly Glu Thr Lys Ala Phe Tyr Pro 485 490 495
gag gag atc tcg tcc atg gtg ctg acc aag atg aag gag atc gcc gag
1536Glu Glu Ile Ser Ser Met Val Leu Thr Lys Met Lys Glu Ile Ala Glu
500 505 510 gcg tac ctg ggc tac ccg gtg acc aac gcg gtg atc acc gtg
ccg gcc 1584Ala Tyr Leu Gly Tyr Pro Val Thr Asn Ala Val Ile Thr Val
Pro Ala 515 520 525 tac ttc aac gac tcg cag cgc cag gcc acc aag gat
gcg ggt gtg atc 1632Tyr Phe Asn Asp Ser Gln Arg Gln Ala Thr Lys Asp
Ala Gly Val Ile 530 535 540 gcg ggg ctc aac gtg ctg cgg atc atc aac
gag ccc acg gcc gcc gcc 1680Ala Gly Leu Asn Val Leu Arg Ile Ile Asn
Glu Pro Thr Ala Ala Ala 545 550 555 560 atc gcc tac ggc ctg gac aga
acg ggc aag ggg gag cgc aac gtg ctc 1728Ile Ala Tyr Gly Leu Asp Arg
Thr Gly Lys Gly Glu Arg Asn Val Leu 565 570 575 atc ttt gac ctg ggc
ggg ggc acc ttc gac gtg tcc atc ctg acg atc 1776Ile Phe Asp Leu Gly
Gly Gly Thr Phe Asp Val Ser Ile Leu Thr Ile 580 585 590 gac gac ggc
atc ttc gag gtg aag gcc acg gcc ggg gac acc cac ctg 1824Asp Asp Gly
Ile Phe Glu Val Lys Ala Thr Ala Gly Asp Thr His Leu 595 600 605 ggt
ggg gag gac ttt gac aac agg ctg gtg aac cac ttc gtg gag gag 1872Gly
Gly Glu Asp Phe Asp Asn Arg Leu Val Asn His Phe Val Glu Glu 610 615
620 ttc aag aga aaa cac aag aag gac atc agc cag aac aag cga gcc gtg
1920Phe Lys Arg Lys His Lys Lys Asp Ile Ser Gln Asn Lys Arg Ala Val
625 630 635 640 agg cgg ctg cgc acc gcc tgc gag agg gcc aag agg acc
ctg tcg tcc 1968Arg Arg Leu Arg Thr Ala Cys Glu Arg Ala Lys Arg Thr
Leu Ser Ser 645 650 655 agc acc cag gcc agc ctg gag atc gac tcc ctg
ttt gag ggc atc gac 2016Ser Thr Gln Ala Ser Leu Glu Ile Asp Ser Leu
Phe Glu Gly Ile Asp 660 665 670 ttc tac acg tcc atc acc agg gcg agg
ttc gag gag ctg tgc tcc gac 2064Phe Tyr Thr Ser Ile Thr Arg Ala Arg
Phe Glu Glu Leu Cys Ser Asp 675 680 685 ctg ttc cga agc acc ctg gag
ccc gtg gag aag gct ctg cgc gac gcc 2112Leu Phe Arg Ser Thr Leu Glu
Pro Val Glu Lys Ala Leu Arg Asp Ala 690 695 700 aag ctg gac aag gcc
cag att cac gac ctg gtc ctg gtc ggg ggc tcc 2160Lys Leu Asp Lys Ala
Gln Ile His Asp Leu Val Leu Val Gly Gly Ser 705 710 715 720 acc cgc
atc ccc aag gtg cag aag ctg ctg cag gac ttc ttc aac ggg 2208Thr Arg
Ile Pro Lys Val Gln Lys Leu Leu Gln Asp Phe Phe Asn Gly 725 730 735
cgc gac ctg aac aag agc atc aac ccc gac gag gct gtg gcc tac ggg
2256Arg Asp Leu Asn Lys Ser Ile Asn Pro Asp Glu Ala Val Ala Tyr Gly
740 745 750 gcg gcg gtg cag gcg gcc atc ctg atg ggg gac aag tcc gag
aac gtg 2304Ala Ala Val Gln Ala Ala Ile Leu Met Gly Asp Lys Ser Glu
Asn Val 755 760 765 cag gac ctg ctg ctg ctg gac gtg gct ccc ctg tcg
ctg ggg ctg gag 2352Gln Asp Leu Leu Leu Leu Asp Val Ala Pro Leu Ser
Leu Gly Leu Glu 770 775 780 acg gcc gga ggc gtg atg act gcc ctg atc
aag cgc aac tcc acc atc 2400Thr Ala Gly Gly Val Met Thr Ala Leu Ile
Lys Arg Asn Ser Thr Ile 785 790 795 800 ccc acc aag cag acg cag atc
ttc acc acc tac tcc gac aac caa ccc 2448Pro Thr Lys Gln Thr Gln Ile
Phe Thr Thr Tyr Ser Asp Asn Gln Pro 805 810 815 ggg gtg ctg atc cag
gtg tac gag ggc gag agg gcc atg acg aaa gac 2496Gly Val Leu Ile Gln
Val Tyr Glu Gly Glu Arg Ala Met Thr Lys Asp 820 825 830 aac aat ctg
ttg ggg cgc ttc gag ctg agc ggc atc cct ccg gcc ccc 2544Asn Asn Leu
Leu Gly Arg Phe Glu Leu Ser Gly Ile Pro Pro Ala Pro 835 840 845 agg
ggc gtg ccc cag atc gag gtg acc ttc gac atc gat gcc aac ggc 2592Arg
Gly Val Pro Gln Ile Glu Val Thr Phe Asp Ile Asp Ala Asn Gly 850 855
860 atc ctg aac gtc acg gcc acg gac aag agc acc ggc aag gcc aac aag
2640Ile Leu Asn Val Thr Ala Thr Asp Lys Ser Thr Gly Lys Ala Asn Lys
865 870 875 880 atc acc atc acc aac gac aag ggc cgc ctg agc aag gag
gag atc gag 2688Ile Thr Ile Thr Asn Asp Lys Gly Arg Leu Ser Lys Glu
Glu Ile Glu 885 890 895 cgc atg gtg cag gag gcg gag aag tac aaa gcg
gag gac gag gtg cag 2736Arg Met Val Gln Glu Ala Glu Lys Tyr Lys Ala
Glu Asp Glu Val Gln 900 905 910 cgc gag agg gtg tca gcc aag aac gcc
ctg gag tcc tac gcc ttc aac 2784Arg Glu Arg Val Ser Ala Lys Asn Ala
Leu Glu Ser Tyr Ala Phe Asn 915 920 925 atg aag agc gcc gtg gag gat
gag ggg ctc aag ggc aag atc agc gag 2832Met Lys Ser Ala Val Glu Asp
Glu Gly Leu Lys Gly Lys Ile Ser Glu 930 935 940 gcg gac aag aag aag
gtt ctg gac aag tgt caa gag gtc atc tcg tgg 2880Ala Asp Lys Lys Lys
Val Leu Asp Lys Cys Gln Glu Val Ile Ser Trp 945 950 955 960 ctg gac
gcc aac acc ttg gcc gag aag gac gag ttt gag cac aag agg 2928Leu Asp
Ala Asn Thr Leu Ala Glu Lys Asp Glu Phe Glu His Lys Arg 965 970 975
aag gag ctg gag cag gtg tgt aac ccc atc atc agc gga ctg tac cag
2976Lys Glu Leu Glu Gln Val Cys Asn Pro Ile Ile Ser Gly Leu Tyr Gln
980 985 990 ggt gcc ggt ggt ccc ggg cct ggc ggc ttc ggg gct cag ggt
ccc aag 3024Gly Ala Gly Gly Pro Gly Pro Gly Gly Phe Gly Ala Gln Gly
Pro Lys 995 1000 1005 gga ggg tct ggg tca ggc cct acc att gag gag
gtg gat tag tctaga 3072Gly Gly Ser Gly Ser Gly Pro Thr Ile Glu Glu
Val Asp 1010 1015 1020 51021PRTArtificial SequenceSynthetic
Construct 5Met Arg Phe Pro Ser Leu Phe Thr Ala Val Leu Phe Ala Ala
Ser Ser 1 5 10 15 Ala Leu Ala Ala Pro Val Asn Thr Thr Thr Glu Asp
Glu Thr Ala Gln 20 25 30 Ile Pro Ala Glu Ala Val Ile Gly Tyr Ser
Asp Leu Glu Gly Asp Phe 35 40 45 Asp Val Ala Val Leu Pro Phe Ser
Asn Ser Thr Asn Asn Gly Leu Leu 50 55 60 Phe Ile Asn Thr Thr Ile
Ala Ser Ile Ala Ala Lys Glu Glu Gly Val 65 70 75 80 Ser Leu Glu Lys
Arg Glu Ala Glu Ala Glu Phe His His His His His 85 90 95 His Ala
Gly Ile His Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu 100 105 110
Ala Val Ser Leu Gly Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Lys 115
120 125 Ser Val Ser Thr Ser Ser Tyr Ser Tyr Met His Trp Tyr Gln Gln
Lys 130 135 140 Pro Gly Gln Pro Pro Lys Leu Leu Ile Lys Tyr Ala Ser
Tyr Leu Glu 145 150 155 160 Ser Gly Val Pro Ala Arg Phe Ser Gly Ser
Gly Ser Gly Thr Asp Phe 165 170 175 Thr Leu Asn Ile His Pro Val Glu
Glu Glu Asp Ala Ala Thr Tyr Tyr 180 185 190 Cys Gln His Ser Arg Glu
Phe Pro Trp Thr Phe Gly Gly Gly Thr Lys 195 200 205 Leu Glu Ile Lys
Arg Ala Asp Ala Ala Pro Gly Gly Gly Gly Ser Gly 210 215 220 Gly Gly
Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser 225 230 235
240 Gly Gly Gly Leu Val Lys Pro Gly Gly Ser Arg Lys Leu Ser Cys Ala
245 250 255 Ala Ser Gly Phe Thr Phe Ser Asn Tyr Gly Met His Trp Val
Arg Gln 260 265 270 Ala Pro Glu Lys Gly Leu Glu Trp Val Ala Tyr Ile
Ser Ser Gly Ser 275 280 285 Ser Thr Ile Tyr Tyr Ala Asp Thr Val Lys
Gly Arg Phe Thr Ile Ser 290 295 300 Arg Asp Asn Ala Lys Asn Thr Leu
Phe Leu Gln Met Thr Ser Leu Arg 305 310 315 320 Ser Glu Asp Thr Ala
Met Arg Gly Leu Leu Leu Asp Tyr Trp Gly Gln 325 330 335 Gly Thr Thr
Leu Thr Val Ser Ser Leu Glu Gln Lys Leu Ile Ser Glu 340 345 350 Glu
Asp Leu Asn Ser Ala Val Asp Ala Lys Thr Thr Ala Pro Ser Val 355 360
365 Tyr Pro Leu Ala Pro Val Leu Glu Ser Ser Gly Ser Met Ala Lys Ala
370 375 380 Ala Ala Ile Gly Ile Asp Leu Gly Thr Thr Tyr Ser Cys Val
Gly Val 385 390 395 400 Phe Gln His Gly Lys Val Glu Ile Ile Ala Asn
Asp Gln Gly Asn Arg 405 410 415 Thr Thr Pro Ser Tyr Val Ala Phe Thr
Asp Thr Glu Arg Leu Ile Gly 420 425 430 Asp Ala Ala Lys Asn Gln Val
Ala Leu Asn Pro Gln Asn Thr Val Phe 435 440 445 Asp Ala Lys Arg Leu
Ile Gly Arg Lys Phe Gly Asp Pro Val Val Gln 450 455 460 Ser Asp Met
Lys His Trp Pro Phe Gln Val Ile Asn Asp Gly Asp Lys 465 470 475 480
Pro Lys Val Gln Val Ser Tyr Lys Gly Glu Thr Lys Ala Phe Tyr Pro 485
490 495 Glu Glu Ile Ser Ser Met Val Leu Thr Lys Met Lys Glu Ile Ala
Glu 500 505 510 Ala Tyr Leu Gly Tyr Pro Val Thr Asn Ala Val Ile Thr
Val Pro Ala 515 520 525 Tyr Phe Asn Asp Ser Gln Arg Gln Ala Thr Lys
Asp Ala Gly Val Ile 530 535 540 Ala Gly Leu Asn
Val Leu Arg Ile Ile Asn Glu Pro Thr Ala Ala Ala 545 550 555 560 Ile
Ala Tyr Gly Leu Asp Arg Thr Gly Lys Gly Glu Arg Asn Val Leu 565 570
575 Ile Phe Asp Leu Gly Gly Gly Thr Phe Asp Val Ser Ile Leu Thr Ile
580 585 590 Asp Asp Gly Ile Phe Glu Val Lys Ala Thr Ala Gly Asp Thr
His Leu 595 600 605 Gly Gly Glu Asp Phe Asp Asn Arg Leu Val Asn His
Phe Val Glu Glu 610 615 620 Phe Lys Arg Lys His Lys Lys Asp Ile Ser
Gln Asn Lys Arg Ala Val 625 630 635 640 Arg Arg Leu Arg Thr Ala Cys
Glu Arg Ala Lys Arg Thr Leu Ser Ser 645 650 655 Ser Thr Gln Ala Ser
Leu Glu Ile Asp Ser Leu Phe Glu Gly Ile Asp 660 665 670 Phe Tyr Thr
Ser Ile Thr Arg Ala Arg Phe Glu Glu Leu Cys Ser Asp 675 680 685 Leu
Phe Arg Ser Thr Leu Glu Pro Val Glu Lys Ala Leu Arg Asp Ala 690 695
700 Lys Leu Asp Lys Ala Gln Ile His Asp Leu Val Leu Val Gly Gly Ser
705 710 715 720 Thr Arg Ile Pro Lys Val Gln Lys Leu Leu Gln Asp Phe
Phe Asn Gly 725 730 735 Arg Asp Leu Asn Lys Ser Ile Asn Pro Asp Glu
Ala Val Ala Tyr Gly 740 745 750 Ala Ala Val Gln Ala Ala Ile Leu Met
Gly Asp Lys Ser Glu Asn Val 755 760 765 Gln Asp Leu Leu Leu Leu Asp
Val Ala Pro Leu Ser Leu Gly Leu Glu 770 775 780 Thr Ala Gly Gly Val
Met Thr Ala Leu Ile Lys Arg Asn Ser Thr Ile 785 790 795 800 Pro Thr
Lys Gln Thr Gln Ile Phe Thr Thr Tyr Ser Asp Asn Gln Pro 805 810 815
Gly Val Leu Ile Gln Val Tyr Glu Gly Glu Arg Ala Met Thr Lys Asp 820
825 830 Asn Asn Leu Leu Gly Arg Phe Glu Leu Ser Gly Ile Pro Pro Ala
Pro 835 840 845 Arg Gly Val Pro Gln Ile Glu Val Thr Phe Asp Ile Asp
Ala Asn Gly 850 855 860 Ile Leu Asn Val Thr Ala Thr Asp Lys Ser Thr
Gly Lys Ala Asn Lys 865 870 875 880 Ile Thr Ile Thr Asn Asp Lys Gly
Arg Leu Ser Lys Glu Glu Ile Glu 885 890 895 Arg Met Val Gln Glu Ala
Glu Lys Tyr Lys Ala Glu Asp Glu Val Gln 900 905 910 Arg Glu Arg Val
Ser Ala Lys Asn Ala Leu Glu Ser Tyr Ala Phe Asn 915 920 925 Met Lys
Ser Ala Val Glu Asp Glu Gly Leu Lys Gly Lys Ile Ser Glu 930 935 940
Ala Asp Lys Lys Lys Val Leu Asp Lys Cys Gln Glu Val Ile Ser Trp 945
950 955 960 Leu Asp Ala Asn Thr Leu Ala Glu Lys Asp Glu Phe Glu His
Lys Arg 965 970 975 Lys Glu Leu Glu Gln Val Cys Asn Pro Ile Ile Ser
Gly Leu Tyr Gln 980 985 990 Gly Ala Gly Gly Pro Gly Pro Gly Gly Phe
Gly Ala Gln Gly Pro Lys 995 1000 1005 Gly Gly Ser Gly Ser Gly Pro
Thr Ile Glu Glu Val Asp 1010 1015 1020 61021PRTArtificial
Sequence3E10 Fv-Hsp70 amino acid sequence for a chimeric protein
comprising a single chain Fv antibody fragment derived from Mus
musculus 3E10 hybridoma and Hsp70 from Homo sapiens joined by mouse
CH1 linker and swivel sequence 6Met Arg Phe Pro Ser Leu Phe Thr Ala
Val Leu Phe Ala Ala Ser Ser 1 5 10 15 Ala Leu Ala Ala Pro Val Asn
Thr Thr Thr Glu Asp Glu Thr Ala Gln 20 25 30 Ile Pro Ala Glu Ala
Val Ile Gly Tyr Ser Asp Leu Glu Gly Asp Phe 35 40 45 Asp Val Ala
Val Leu Pro Phe Ser Asn Ser Thr Asn Asn Gly Leu Leu 50 55 60 Phe
Ile Asn Thr Thr Ile Ala Ser Ile Ala Ala Lys Glu Glu Gly Val 65 70
75 80 Ser Leu Glu Lys Arg Glu Ala Glu Ala Glu Phe His His His His
His 85 90 95 His Ala Gly Ile His Asp Ile Val Leu Thr Gln Ser Pro
Ala Ser Leu 100 105 110 Ala Val Ser Leu Gly Gln Arg Ala Thr Ile Ser
Cys Arg Ala Ser Lys 115 120 125 Ser Val Ser Thr Ser Ser Tyr Ser Tyr
Met His Trp Tyr Gln Gln Lys 130 135 140 Pro Gly Gln Pro Pro Lys Leu
Leu Ile Lys Tyr Ala Ser Tyr Leu Glu 145 150 155 160 Ser Gly Val Pro
Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe 165 170 175 Thr Leu
Asn Ile His Pro Val Glu Glu Glu Asp Ala Ala Thr Tyr Tyr 180 185 190
Cys Gln His Ser Arg Glu Phe Pro Trp Thr Phe Gly Gly Gly Thr Lys 195
200 205 Leu Glu Ile Lys Arg Ala Asp Ala Ala Pro Gly Gly Gly Gly Ser
Gly 210 215 220 Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu
Val Glu Ser 225 230 235 240 Gly Gly Gly Leu Val Lys Pro Gly Gly Ser
Arg Lys Leu Ser Cys Ala 245 250 255 Ala Ser Gly Phe Thr Phe Ser Asn
Tyr Gly Met His Trp Val Arg Gln 260 265 270 Ala Pro Glu Lys Gly Leu
Glu Trp Val Ala Tyr Ile Ser Ser Gly Ser 275 280 285 Ser Thr Ile Tyr
Tyr Ala Asp Thr Val Lys Gly Arg Phe Thr Ile Ser 290 295 300 Arg Asp
Asn Ala Lys Asn Thr Leu Phe Leu Gln Met Thr Ser Leu Arg 305 310 315
320 Ser Glu Asp Thr Ala Met Arg Gly Leu Leu Leu Asp Tyr Trp Gly Gln
325 330 335 Gly Thr Thr Leu Thr Val Ser Ser Leu Glu Gln Lys Leu Ile
Ser Glu 340 345 350 Glu Asp Leu Asn Ser Ala Val Asp Ala Lys Thr Thr
Ala Pro Ser Val 355 360 365 Tyr Pro Leu Ala Pro Val Leu Glu Ser Ser
Gly Ser Met Ala Lys Ala 370 375 380 Ala Ala Ile Gly Ile Asp Leu Gly
Thr Thr Tyr Ser Cys Val Gly Val 385 390 395 400 Phe Gln His Gly Lys
Val Glu Ile Ile Ala Asn Asp Gln Gly Asn Arg 405 410 415 Thr Thr Pro
Ser Tyr Val Ala Phe Thr Asp Thr Glu Arg Leu Ile Gly 420 425 430 Asp
Ala Ala Lys Asn Gln Val Ala Leu Asn Pro Gln Asn Thr Val Phe 435 440
445 Asp Ala Lys Arg Leu Ile Gly Arg Lys Phe Gly Asp Pro Val Val Gln
450 455 460 Ser Asp Met Lys His Trp Pro Phe Gln Val Ile Asn Asp Gly
Asp Lys 465 470 475 480 Pro Lys Val Gln Val Ser Tyr Lys Gly Glu Thr
Lys Ala Phe Tyr Pro 485 490 495 Glu Glu Ile Ser Ser Met Val Leu Thr
Lys Met Lys Glu Ile Ala Glu 500 505 510 Ala Tyr Leu Gly Tyr Pro Val
Thr Asn Ala Val Ile Thr Val Pro Ala 515 520 525 Tyr Phe Asn Asp Ser
Gln Arg Gln Ala Thr Lys Asp Ala Gly Val Ile 530 535 540 Ala Gly Leu
Asn Val Leu Arg Ile Ile Asn Glu Pro Thr Ala Ala Ala 545 550 555 560
Ile Ala Tyr Gly Leu Asp Arg Thr Gly Lys Gly Glu Arg Asn Val Leu 565
570 575 Ile Phe Asp Leu Gly Gly Gly Thr Phe Asp Val Ser Ile Leu Thr
Ile 580 585 590 Asp Asp Gly Ile Phe Glu Val Lys Ala Thr Ala Gly Asp
Thr His Leu 595 600 605 Gly Gly Glu Asp Phe Asp Asn Arg Leu Val Asn
His Phe Val Glu Glu 610 615 620 Phe Lys Arg Lys His Lys Lys Asp Ile
Ser Gln Asn Lys Arg Ala Val 625 630 635 640 Arg Arg Leu Arg Thr Ala
Cys Glu Arg Ala Lys Arg Thr Leu Ser Ser 645 650 655 Ser Thr Gln Ala
Ser Leu Glu Ile Asp Ser Leu Phe Glu Gly Ile Asp 660 665 670 Phe Tyr
Thr Ser Ile Thr Arg Ala Arg Phe Glu Glu Leu Cys Ser Asp 675 680 685
Leu Phe Arg Ser Thr Leu Glu Pro Val Glu Lys Ala Leu Arg Asp Ala 690
695 700 Lys Leu Asp Lys Ala Gln Ile His Asp Leu Val Leu Val Gly Gly
Ser 705 710 715 720 Thr Arg Ile Pro Lys Val Gln Lys Leu Leu Gln Asp
Phe Phe Asn Gly 725 730 735 Arg Asp Leu Asn Lys Ser Ile Asn Pro Asp
Glu Ala Val Ala Tyr Gly 740 745 750 Ala Ala Val Gln Ala Ala Ile Leu
Met Gly Asp Lys Ser Glu Asn Val 755 760 765 Gln Asp Leu Leu Leu Leu
Asp Val Ala Pro Leu Ser Leu Gly Leu Glu 770 775 780 Thr Ala Gly Gly
Val Met Thr Ala Leu Ile Lys Arg Asn Ser Thr Ile 785 790 795 800 Pro
Thr Lys Gln Thr Gln Ile Phe Thr Thr Tyr Ser Asp Asn Gln Pro 805 810
815 Gly Val Leu Ile Gln Val Tyr Glu Gly Glu Arg Ala Met Thr Lys Asp
820 825 830 Asn Asn Leu Leu Gly Arg Phe Glu Leu Ser Gly Ile Pro Pro
Ala Pro 835 840 845 Arg Gly Val Pro Gln Ile Glu Val Thr Phe Asp Ile
Asp Ala Asn Gly 850 855 860 Ile Leu Asn Val Thr Ala Thr Asp Lys Ser
Thr Gly Lys Ala Asn Lys 865 870 875 880 Ile Thr Ile Thr Asn Asp Lys
Gly Arg Leu Ser Lys Glu Glu Ile Glu 885 890 895 Arg Met Val Gln Glu
Ala Glu Lys Tyr Lys Ala Glu Asp Glu Val Gln 900 905 910 Arg Glu Arg
Val Ser Ala Lys Asn Ala Leu Glu Ser Tyr Ala Phe Asn 915 920 925 Met
Lys Ser Ala Val Glu Asp Glu Gly Leu Lys Gly Lys Ile Ser Glu 930 935
940 Ala Asp Lys Lys Lys Val Leu Asp Lys Cys Gln Glu Val Ile Ser Trp
945 950 955 960 Leu Asp Ala Asn Thr Leu Ala Glu Lys Asp Glu Phe Glu
His Lys Arg 965 970 975 Lys Glu Leu Glu Gln Val Cys Asn Pro Ile Ile
Ser Gly Leu Tyr Gln 980 985 990 Gly Ala Gly Gly Pro Gly Pro Gly Gly
Phe Gly Ala Gln Gly Pro Lys 995 1000 1005 Gly Gly Ser Gly Ser Gly
Pro Thr Ile Glu Glu Val Asp 1010 1015 1020 73111DNAArtificial
Sequence3E10 Fv-GRP78 nucleotide sequence encoding a chimeric
protein comprising a single chain Fv antibody fragment derived from
Mus musculus 3E10 hybridoma and GRP78 from Homo sapiens joined by
mouse CH1 linker and swivel sequence 7atg aga ttt cct tca ctt ttt
act gct gtt tta ttc gca gca tcc tcc 48Met Arg Phe Pro Ser Leu Phe
Thr Ala Val Leu Phe Ala Ala Ser Ser 1 5 10 15 gca tta gct gct cca
gtc aac act aca aca gaa gat gaa acg gca caa 96Ala Leu Ala Ala Pro
Val Asn Thr Thr Thr Glu Asp Glu Thr Ala Gln 20 25 30 att ccg gct
gaa gct gtc atc ggt tac tca gat tta gaa ggg gat ttc 144Ile Pro Ala
Glu Ala Val Ile Gly Tyr Ser Asp Leu Glu Gly Asp Phe 35 40 45 gat
gtt gct gtt ttg cca ttt tcc aac agc aca aat aac ggg tta ttg 192Asp
Val Ala Val Leu Pro Phe Ser Asn Ser Thr Asn Asn Gly Leu Leu 50 55
60 ttt ata aat act act att gcc agc att gct gct aaa gaa gaa ggg gta
240Phe Ile Asn Thr Thr Ile Ala Ser Ile Ala Ala Lys Glu Glu Gly Val
65 70 75 80 tct ctc gag aaa aga gag gct gaa gct gaa ttc cat cac cat
cac cat 288Ser Leu Glu Lys Arg Glu Ala Glu Ala Glu Phe His His His
His His 85 90 95 cac gca ggg att cac gac att gtc ctg aca cag tct
cct gct tcc tta 336His Ala Gly Ile His Asp Ile Val Leu Thr Gln Ser
Pro Ala Ser Leu 100 105 110 gct gta tct ctg ggg cag agg gcc acc atc
tcc tgc agg gcc agc aaa 384Ala Val Ser Leu Gly Gln Arg Ala Thr Ile
Ser Cys Arg Ala Ser Lys 115 120 125 agt gtc agt aca tct agc tat agt
tac atg cac tgg tac caa cag aaa 432Ser Val Ser Thr Ser Ser Tyr Ser
Tyr Met His Trp Tyr Gln Gln Lys 130 135 140 cca gga cag cca ccc aaa
ctc ctc atc aag tat gca tcc tac cta gaa 480Pro Gly Gln Pro Pro Lys
Leu Leu Ile Lys Tyr Ala Ser Tyr Leu Glu 145 150 155 160 tct ggg gtt
cct gcc agg ttc agt ggc agt ggg tct ggg aca gac ttc 528Ser Gly Val
Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe 165 170 175 acc
ctc aac atc cat cct gtg gag gag gag gat gct gca aca tat tac 576Thr
Leu Asn Ile His Pro Val Glu Glu Glu Asp Ala Ala Thr Tyr Tyr 180 185
190 tgt cag cac agt agg gag ttt ccg tgg acg ttc ggt gga ggc acc aag
624Cys Gln His Ser Arg Glu Phe Pro Trp Thr Phe Gly Gly Gly Thr Lys
195 200 205 ctg gaa atc aaa cgg gct gat gct gca ccc ggg ggt ggc ggt
tct ggc 672Leu Glu Ile Lys Arg Ala Asp Ala Ala Pro Gly Gly Gly Gly
Ser Gly 210 215 220 ggt ggc ggt tct gga ggc ggt ggc tct gag gtg cag
ctg gtg gag tct 720Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln
Leu Val Glu Ser 225 230 235 240 ggg gga ggc tta gtg aag cct gga ggg
tcc cgg aaa ctc tcc tgt gca 768Gly Gly Gly Leu Val Lys Pro Gly Gly
Ser Arg Lys Leu Ser Cys Ala 245 250 255 gcc tct gga ttc act ttc agt
aac tat gga atg cac tgg gtc cgt cag 816Ala Ser Gly Phe Thr Phe Ser
Asn Tyr Gly Met His Trp Val Arg Gln 260 265 270 gct cca gag aag ggg
ctg gag tgg gtt gca tac att agt agt ggc agt 864Ala Pro Glu Lys Gly
Leu Glu Trp Val Ala Tyr Ile Ser Ser Gly Ser 275 280 285 agt acc atc
tac tat gca gac aca gtg aag ggc cga ttc acc atc tcc 912Ser Thr Ile
Tyr Tyr Ala Asp Thr Val Lys Gly Arg Phe Thr Ile Ser 290 295 300 aga
gac aat gcc aag aac acc ctg ttc ctg caa atg acc agt cta agg 960Arg
Asp Asn Ala Lys Asn Thr Leu Phe Leu Gln Met Thr Ser Leu Arg 305 310
315 320 tct gag gac aca gcc atg cgg ggg tta cta ctt gac tac tgg ggc
caa 1008Ser Glu Asp Thr Ala Met Arg Gly Leu Leu Leu Asp Tyr Trp Gly
Gln 325 330 335 ggc acc act ctc aca gtc tcc tca cta gaa caa aaa ctc
atc tca gaa 1056Gly Thr Thr Leu Thr Val Ser Ser Leu Glu Gln Lys Leu
Ile Ser Glu 340 345 350 gag gat ctg aat agc gcc gtc gac gcc aaa aca
aca gcc cca tcg gtc 1104Glu Asp Leu Asn Ser Ala Val Asp Ala Lys Thr
Thr Ala Pro Ser Val 355 360 365 tat cca ctg gcc cct gta ctg gag tct
tcc gga tcc atg aag ctc tcc 1152Tyr Pro Leu Ala Pro Val Leu Glu Ser
Ser Gly Ser Met Lys Leu Ser 370 375 380 ctg gtg gcc gcg atg ctg ctg
ctg ctc agc gcg gcg cgg gcc gag gag 1200Leu Val Ala Ala Met Leu Leu
Leu Leu Ser Ala Ala Arg Ala Glu Glu 385 390 395 400 gag gac aag aag
gag gac gtg ggc acg gtg gtc ggc atc gac ctg ggg 1248Glu Asp Lys Lys
Glu Asp Val Gly Thr Val Val Gly Ile Asp Leu Gly 405 410 415 acc acc
tac tcc tgc gtc ggc gtg ttc aag aac ggc cgc gtg gag atc 1296Thr Thr
Tyr Ser Cys Val Gly Val Phe Lys Asn Gly Arg Val Glu Ile 420
425 430 atc gcc aac gat cag ggc aac cgc atc acg ccg tcc tat gtc gcc
ttc 1344Ile Ala Asn Asp Gln Gly Asn Arg Ile Thr Pro Ser Tyr Val Ala
Phe 435 440 445 act cct gaa ggg gaa cgt ctg att ggc gat gcc gcc aag
aac cag ctc 1392Thr Pro Glu Gly Glu Arg Leu Ile Gly Asp Ala Ala Lys
Asn Gln Leu 450 455 460 acc tcc aac ccc gag aac acg gtc ttt gac gcc
aag cgg ctc atc ggc 1440Thr Ser Asn Pro Glu Asn Thr Val Phe Asp Ala
Lys Arg Leu Ile Gly 465 470 475 480 cgc acg tgg aat gac ccg tct gtg
cag cag gac atc aag ttc ttg ccg 1488Arg Thr Trp Asn Asp Pro Ser Val
Gln Gln Asp Ile Lys Phe Leu Pro 485 490 495 ttc aag gtg gtt gaa aag
aaa act aaa cca tac att caa gtt gat att 1536Phe Lys Val Val Glu Lys
Lys Thr Lys Pro Tyr Ile Gln Val Asp Ile 500 505 510 gga ggt ggg caa
aca aag aca ttt gct cct gaa gaa att tct gcc atg 1584Gly Gly Gly Gln
Thr Lys Thr Phe Ala Pro Glu Glu Ile Ser Ala Met 515 520 525 gtt ctc
act aaa atg aaa gaa acc gct gag gct tat ttg gga aag aag 1632Val Leu
Thr Lys Met Lys Glu Thr Ala Glu Ala Tyr Leu Gly Lys Lys 530 535 540
gtt acc cat gca gtt gtt act gta cca gcc tat ttt aat gat gcc caa
1680Val Thr His Ala Val Val Thr Val Pro Ala Tyr Phe Asn Asp Ala Gln
545 550 555 560 cgc caa gca acc aaa gac gct gga act att gct ggc cta
aat gtt atg 1728Arg Gln Ala Thr Lys Asp Ala Gly Thr Ile Ala Gly Leu
Asn Val Met 565 570 575 agg atc atc aac gag cct acg gca gct gct att
gct tat ggc ctg gat 1776Arg Ile Ile Asn Glu Pro Thr Ala Ala Ala Ile
Ala Tyr Gly Leu Asp 580 585 590 aag agg gag ggg gag aag aac atc ctg
gtg ttt gac ctg ggt ggc gga 1824Lys Arg Glu Gly Glu Lys Asn Ile Leu
Val Phe Asp Leu Gly Gly Gly 595 600 605 acc ttc gat gtg tct ctt ctc
acc att gac aat ggt gtc ttc gaa gtt 1872Thr Phe Asp Val Ser Leu Leu
Thr Ile Asp Asn Gly Val Phe Glu Val 610 615 620 gtg gcc act aat gga
gat act cat ctg ggt gga gaa gac ttt gac cag 1920Val Ala Thr Asn Gly
Asp Thr His Leu Gly Gly Glu Asp Phe Asp Gln 625 630 635 640 cgt gtc
atg gaa cac ttc atc aaa ctg tac aaa aag aag acg ggc aaa 1968Arg Val
Met Glu His Phe Ile Lys Leu Tyr Lys Lys Lys Thr Gly Lys 645 650 655
gat gtc agg aaa gac aat aga gct gtg cag aaa ctc cgg cgc gag gta
2016Asp Val Arg Lys Asp Asn Arg Ala Val Gln Lys Leu Arg Arg Glu Val
660 665 670 gaa aag gcc aaa cgg gcc ctg tct tct cag cat caa gca aga
att gaa 2064Glu Lys Ala Lys Arg Ala Leu Ser Ser Gln His Gln Ala Arg
Ile Glu 675 680 685 att gag tcc ttc tat gaa gga gaa gac ttt tct gag
acc ctg act cgg 2112Ile Glu Ser Phe Tyr Glu Gly Glu Asp Phe Ser Glu
Thr Leu Thr Arg 690 695 700 gcc aaa ttt gaa gag ctc aac atg gat ctg
ttc cgg tct act atg aag 2160Ala Lys Phe Glu Glu Leu Asn Met Asp Leu
Phe Arg Ser Thr Met Lys 705 710 715 720 ccc gtc cag aaa gtg ttg gaa
gat tct gat ttg aag aag tct gat att 2208Pro Val Gln Lys Val Leu Glu
Asp Ser Asp Leu Lys Lys Ser Asp Ile 725 730 735 gat gaa att gtt ctt
gtt ggt ggc tcg act cga att cca aag att cag 2256Asp Glu Ile Val Leu
Val Gly Gly Ser Thr Arg Ile Pro Lys Ile Gln 740 745 750 caa ctg gtt
aaa gag ttc ttc aat ggc aag gaa cca tcc cgt ggc ata 2304Gln Leu Val
Lys Glu Phe Phe Asn Gly Lys Glu Pro Ser Arg Gly Ile 755 760 765 aac
cca gat gaa gct gta gcg tat ggt gct gct gtc cag gct ggt gtg 2352Asn
Pro Asp Glu Ala Val Ala Tyr Gly Ala Ala Val Gln Ala Gly Val 770 775
780 ctc tct ggt gat caa gat aca ggt gac ctg gta ctg ctt gat gta tgt
2400Leu Ser Gly Asp Gln Asp Thr Gly Asp Leu Val Leu Leu Asp Val Cys
785 790 795 800 ccc ctt aca ctt ggt att gaa act gtg gga ggt gtc atg
acc aaa ctg 2448Pro Leu Thr Leu Gly Ile Glu Thr Val Gly Gly Val Met
Thr Lys Leu 805 810 815 att cca agg aac aca gtg gtg cct acc aag aag
tct cag atc ttt tct 2496Ile Pro Arg Asn Thr Val Val Pro Thr Lys Lys
Ser Gln Ile Phe Ser 820 825 830 aca gct tct gat aat caa cca act gtt
aca atc aag gtc tat gaa ggt 2544Thr Ala Ser Asp Asn Gln Pro Thr Val
Thr Ile Lys Val Tyr Glu Gly 835 840 845 gaa aga ccc ctg aca aaa gac
aat cat ctt ctg ggt aca ttt gat ctg 2592Glu Arg Pro Leu Thr Lys Asp
Asn His Leu Leu Gly Thr Phe Asp Leu 850 855 860 act gga att cct cct
gct cct cgt ggg gtc cca cag att gaa gtc acc 2640Thr Gly Ile Pro Pro
Ala Pro Arg Gly Val Pro Gln Ile Glu Val Thr 865 870 875 880 ttt gag
ata gat gtg aat ggt att ctt cga gtg aca gct gaa gac aag 2688Phe Glu
Ile Asp Val Asn Gly Ile Leu Arg Val Thr Ala Glu Asp Lys 885 890 895
ggt aca ggg aac aaa aat aag atc aca atc acc aat gac cag aat cgc
2736Gly Thr Gly Asn Lys Asn Lys Ile Thr Ile Thr Asn Asp Gln Asn Arg
900 905 910 ctg aca cct gaa gaa atc gaa agg atg gtt aat gat gct gag
aag ttt 2784Leu Thr Pro Glu Glu Ile Glu Arg Met Val Asn Asp Ala Glu
Lys Phe 915 920 925 gct gag gaa gac aaa aag ctc aag gag cgc att gat
act aga aat gag 2832Ala Glu Glu Asp Lys Lys Leu Lys Glu Arg Ile Asp
Thr Arg Asn Glu 930 935 940 ttg gaa agc tat gcc tat tct cta aag aat
cag att gga gat aaa gaa 2880Leu Glu Ser Tyr Ala Tyr Ser Leu Lys Asn
Gln Ile Gly Asp Lys Glu 945 950 955 960 aag ctg gga ggt aaa ctt tcc
tct gaa gat aag gag acc atg gaa aaa 2928Lys Leu Gly Gly Lys Leu Ser
Ser Glu Asp Lys Glu Thr Met Glu Lys 965 970 975 gct gta gaa gaa aag
att gaa tgg ctg gaa agc cac caa gat gct gac 2976Ala Val Glu Glu Lys
Ile Glu Trp Leu Glu Ser His Gln Asp Ala Asp 980 985 990 att gaa gac
ttc aaa gct aag aag aag gaa ctg gaa gaa att gtt caa 3024Ile Glu Asp
Phe Lys Ala Lys Lys Lys Glu Leu Glu Glu Ile Val Gln 995 1000 1005
cca att atc agc aaa ctc tat gga agt gca ggc cct ccc cca act 3069Pro
Ile Ile Ser Lys Leu Tyr Gly Ser Ala Gly Pro Pro Pro Thr 1010 1015
1020 ggt gaa gag gat aca gca gaa aaa gat gag ttg tag tctaga 3111Gly
Glu Glu Asp Thr Ala Glu Lys Asp Glu Leu 1025 1030
81034PRTArtificial SequenceSynthetic Construct 8Met Arg Phe Pro Ser
Leu Phe Thr Ala Val Leu Phe Ala Ala Ser Ser 1 5 10 15 Ala Leu Ala
Ala Pro Val Asn Thr Thr Thr Glu Asp Glu Thr Ala Gln 20 25 30 Ile
Pro Ala Glu Ala Val Ile Gly Tyr Ser Asp Leu Glu Gly Asp Phe 35 40
45 Asp Val Ala Val Leu Pro Phe Ser Asn Ser Thr Asn Asn Gly Leu Leu
50 55 60 Phe Ile Asn Thr Thr Ile Ala Ser Ile Ala Ala Lys Glu Glu
Gly Val 65 70 75 80 Ser Leu Glu Lys Arg Glu Ala Glu Ala Glu Phe His
His His His His 85 90 95 His Ala Gly Ile His Asp Ile Val Leu Thr
Gln Ser Pro Ala Ser Leu 100 105 110 Ala Val Ser Leu Gly Gln Arg Ala
Thr Ile Ser Cys Arg Ala Ser Lys 115 120 125 Ser Val Ser Thr Ser Ser
Tyr Ser Tyr Met His Trp Tyr Gln Gln Lys 130 135 140 Pro Gly Gln Pro
Pro Lys Leu Leu Ile Lys Tyr Ala Ser Tyr Leu Glu 145 150 155 160 Ser
Gly Val Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe 165 170
175 Thr Leu Asn Ile His Pro Val Glu Glu Glu Asp Ala Ala Thr Tyr Tyr
180 185 190 Cys Gln His Ser Arg Glu Phe Pro Trp Thr Phe Gly Gly Gly
Thr Lys 195 200 205 Leu Glu Ile Lys Arg Ala Asp Ala Ala Pro Gly Gly
Gly Gly Ser Gly 210 215 220 Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu
Val Gln Leu Val Glu Ser 225 230 235 240 Gly Gly Gly Leu Val Lys Pro
Gly Gly Ser Arg Lys Leu Ser Cys Ala 245 250 255 Ala Ser Gly Phe Thr
Phe Ser Asn Tyr Gly Met His Trp Val Arg Gln 260 265 270 Ala Pro Glu
Lys Gly Leu Glu Trp Val Ala Tyr Ile Ser Ser Gly Ser 275 280 285 Ser
Thr Ile Tyr Tyr Ala Asp Thr Val Lys Gly Arg Phe Thr Ile Ser 290 295
300 Arg Asp Asn Ala Lys Asn Thr Leu Phe Leu Gln Met Thr Ser Leu Arg
305 310 315 320 Ser Glu Asp Thr Ala Met Arg Gly Leu Leu Leu Asp Tyr
Trp Gly Gln 325 330 335 Gly Thr Thr Leu Thr Val Ser Ser Leu Glu Gln
Lys Leu Ile Ser Glu 340 345 350 Glu Asp Leu Asn Ser Ala Val Asp Ala
Lys Thr Thr Ala Pro Ser Val 355 360 365 Tyr Pro Leu Ala Pro Val Leu
Glu Ser Ser Gly Ser Met Lys Leu Ser 370 375 380 Leu Val Ala Ala Met
Leu Leu Leu Leu Ser Ala Ala Arg Ala Glu Glu 385 390 395 400 Glu Asp
Lys Lys Glu Asp Val Gly Thr Val Val Gly Ile Asp Leu Gly 405 410 415
Thr Thr Tyr Ser Cys Val Gly Val Phe Lys Asn Gly Arg Val Glu Ile 420
425 430 Ile Ala Asn Asp Gln Gly Asn Arg Ile Thr Pro Ser Tyr Val Ala
Phe 435 440 445 Thr Pro Glu Gly Glu Arg Leu Ile Gly Asp Ala Ala Lys
Asn Gln Leu 450 455 460 Thr Ser Asn Pro Glu Asn Thr Val Phe Asp Ala
Lys Arg Leu Ile Gly 465 470 475 480 Arg Thr Trp Asn Asp Pro Ser Val
Gln Gln Asp Ile Lys Phe Leu Pro 485 490 495 Phe Lys Val Val Glu Lys
Lys Thr Lys Pro Tyr Ile Gln Val Asp Ile 500 505 510 Gly Gly Gly Gln
Thr Lys Thr Phe Ala Pro Glu Glu Ile Ser Ala Met 515 520 525 Val Leu
Thr Lys Met Lys Glu Thr Ala Glu Ala Tyr Leu Gly Lys Lys 530 535 540
Val Thr His Ala Val Val Thr Val Pro Ala Tyr Phe Asn Asp Ala Gln 545
550 555 560 Arg Gln Ala Thr Lys Asp Ala Gly Thr Ile Ala Gly Leu Asn
Val Met 565 570 575 Arg Ile Ile Asn Glu Pro Thr Ala Ala Ala Ile Ala
Tyr Gly Leu Asp 580 585 590 Lys Arg Glu Gly Glu Lys Asn Ile Leu Val
Phe Asp Leu Gly Gly Gly 595 600 605 Thr Phe Asp Val Ser Leu Leu Thr
Ile Asp Asn Gly Val Phe Glu Val 610 615 620 Val Ala Thr Asn Gly Asp
Thr His Leu Gly Gly Glu Asp Phe Asp Gln 625 630 635 640 Arg Val Met
Glu His Phe Ile Lys Leu Tyr Lys Lys Lys Thr Gly Lys 645 650 655 Asp
Val Arg Lys Asp Asn Arg Ala Val Gln Lys Leu Arg Arg Glu Val 660 665
670 Glu Lys Ala Lys Arg Ala Leu Ser Ser Gln His Gln Ala Arg Ile Glu
675 680 685 Ile Glu Ser Phe Tyr Glu Gly Glu Asp Phe Ser Glu Thr Leu
Thr Arg 690 695 700 Ala Lys Phe Glu Glu Leu Asn Met Asp Leu Phe Arg
Ser Thr Met Lys 705 710 715 720 Pro Val Gln Lys Val Leu Glu Asp Ser
Asp Leu Lys Lys Ser Asp Ile 725 730 735 Asp Glu Ile Val Leu Val Gly
Gly Ser Thr Arg Ile Pro Lys Ile Gln 740 745 750 Gln Leu Val Lys Glu
Phe Phe Asn Gly Lys Glu Pro Ser Arg Gly Ile 755 760 765 Asn Pro Asp
Glu Ala Val Ala Tyr Gly Ala Ala Val Gln Ala Gly Val 770 775 780 Leu
Ser Gly Asp Gln Asp Thr Gly Asp Leu Val Leu Leu Asp Val Cys 785 790
795 800 Pro Leu Thr Leu Gly Ile Glu Thr Val Gly Gly Val Met Thr Lys
Leu 805 810 815 Ile Pro Arg Asn Thr Val Val Pro Thr Lys Lys Ser Gln
Ile Phe Ser 820 825 830 Thr Ala Ser Asp Asn Gln Pro Thr Val Thr Ile
Lys Val Tyr Glu Gly 835 840 845 Glu Arg Pro Leu Thr Lys Asp Asn His
Leu Leu Gly Thr Phe Asp Leu 850 855 860 Thr Gly Ile Pro Pro Ala Pro
Arg Gly Val Pro Gln Ile Glu Val Thr 865 870 875 880 Phe Glu Ile Asp
Val Asn Gly Ile Leu Arg Val Thr Ala Glu Asp Lys 885 890 895 Gly Thr
Gly Asn Lys Asn Lys Ile Thr Ile Thr Asn Asp Gln Asn Arg 900 905 910
Leu Thr Pro Glu Glu Ile Glu Arg Met Val Asn Asp Ala Glu Lys Phe 915
920 925 Ala Glu Glu Asp Lys Lys Leu Lys Glu Arg Ile Asp Thr Arg Asn
Glu 930 935 940 Leu Glu Ser Tyr Ala Tyr Ser Leu Lys Asn Gln Ile Gly
Asp Lys Glu 945 950 955 960 Lys Leu Gly Gly Lys Leu Ser Ser Glu Asp
Lys Glu Thr Met Glu Lys 965 970 975 Ala Val Glu Glu Lys Ile Glu Trp
Leu Glu Ser His Gln Asp Ala Asp 980 985 990 Ile Glu Asp Phe Lys Ala
Lys Lys Lys Glu Leu Glu Glu Ile Val Gln 995 1000 1005 Pro Ile Ile
Ser Lys Leu Tyr Gly Ser Ala Gly Pro Pro Pro Thr 1010 1015 1020 Gly
Glu Glu Asp Thr Ala Glu Lys Asp Glu Leu 1025 1030
91034PRTArtificial Sequence3E10 Fv-GRP78 amino acid sequence for a
chimeric protein comprising a single chain Fv antibody fragment
derived from Mus musculus 3E10 hybridoma and GRP78 from Homo
sapiens joined by mouse CH1 linker and swivel sequence 9Met Arg Phe
Pro Ser Leu Phe Thr Ala Val Leu Phe Ala Ala Ser Ser 1 5 10 15 Ala
Leu Ala Ala Pro Val Asn Thr Thr Thr Glu Asp Glu Thr Ala Gln 20 25
30 Ile Pro Ala Glu Ala Val Ile Gly Tyr Ser Asp Leu Glu Gly Asp Phe
35 40 45 Asp Val Ala Val Leu Pro Phe Ser Asn Ser Thr Asn Asn Gly
Leu Leu 50 55 60 Phe Ile Asn Thr Thr Ile Ala Ser Ile Ala Ala Lys
Glu Glu Gly Val 65 70 75 80 Ser Leu Glu Lys Arg Glu Ala Glu Ala Glu
Phe His His His His His 85 90 95 His Ala Gly Ile His Asp Ile Val
Leu Thr Gln Ser Pro Ala Ser Leu 100 105 110 Ala Val Ser Leu Gly Gln
Arg Ala Thr Ile Ser Cys Arg Ala Ser Lys 115 120 125 Ser Val Ser Thr
Ser Ser Tyr Ser Tyr Met His Trp Tyr Gln Gln Lys 130 135 140 Pro Gly
Gln Pro Pro Lys Leu Leu Ile Lys Tyr Ala Ser Tyr Leu Glu 145 150 155
160 Ser Gly Val Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe
165 170 175 Thr Leu Asn
Ile His Pro Val Glu Glu Glu Asp Ala Ala Thr Tyr Tyr 180 185 190 Cys
Gln His Ser Arg Glu Phe Pro Trp Thr Phe Gly Gly Gly Thr Lys 195 200
205 Leu Glu Ile Lys Arg Ala Asp Ala Ala Pro Gly Gly Gly Gly Ser Gly
210 215 220 Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val
Glu Ser 225 230 235 240 Gly Gly Gly Leu Val Lys Pro Gly Gly Ser Arg
Lys Leu Ser Cys Ala 245 250 255 Ala Ser Gly Phe Thr Phe Ser Asn Tyr
Gly Met His Trp Val Arg Gln 260 265 270 Ala Pro Glu Lys Gly Leu Glu
Trp Val Ala Tyr Ile Ser Ser Gly Ser 275 280 285 Ser Thr Ile Tyr Tyr
Ala Asp Thr Val Lys Gly Arg Phe Thr Ile Ser 290 295 300 Arg Asp Asn
Ala Lys Asn Thr Leu Phe Leu Gln Met Thr Ser Leu Arg 305 310 315 320
Ser Glu Asp Thr Ala Met Arg Gly Leu Leu Leu Asp Tyr Trp Gly Gln 325
330 335 Gly Thr Thr Leu Thr Val Ser Ser Leu Glu Gln Lys Leu Ile Ser
Glu 340 345 350 Glu Asp Leu Asn Ser Ala Val Asp Ala Lys Thr Thr Ala
Pro Ser Val 355 360 365 Tyr Pro Leu Ala Pro Val Leu Glu Ser Ser Gly
Ser Met Lys Leu Ser 370 375 380 Leu Val Ala Ala Met Leu Leu Leu Leu
Ser Ala Ala Arg Ala Glu Glu 385 390 395 400 Glu Asp Lys Lys Glu Asp
Val Gly Thr Val Val Gly Ile Asp Leu Gly 405 410 415 Thr Thr Tyr Ser
Cys Val Gly Val Phe Lys Asn Gly Arg Val Glu Ile 420 425 430 Ile Ala
Asn Asp Gln Gly Asn Arg Ile Thr Pro Ser Tyr Val Ala Phe 435 440 445
Thr Pro Glu Gly Glu Arg Leu Ile Gly Asp Ala Ala Lys Asn Gln Leu 450
455 460 Thr Ser Asn Pro Glu Asn Thr Val Phe Asp Ala Lys Arg Leu Ile
Gly 465 470 475 480 Arg Thr Trp Asn Asp Pro Ser Val Gln Gln Asp Ile
Lys Phe Leu Pro 485 490 495 Phe Lys Val Val Glu Lys Lys Thr Lys Pro
Tyr Ile Gln Val Asp Ile 500 505 510 Gly Gly Gly Gln Thr Lys Thr Phe
Ala Pro Glu Glu Ile Ser Ala Met 515 520 525 Val Leu Thr Lys Met Lys
Glu Thr Ala Glu Ala Tyr Leu Gly Lys Lys 530 535 540 Val Thr His Ala
Val Val Thr Val Pro Ala Tyr Phe Asn Asp Ala Gln 545 550 555 560 Arg
Gln Ala Thr Lys Asp Ala Gly Thr Ile Ala Gly Leu Asn Val Met 565 570
575 Arg Ile Ile Asn Glu Pro Thr Ala Ala Ala Ile Ala Tyr Gly Leu Asp
580 585 590 Lys Arg Glu Gly Glu Lys Asn Ile Leu Val Phe Asp Leu Gly
Gly Gly 595 600 605 Thr Phe Asp Val Ser Leu Leu Thr Ile Asp Asn Gly
Val Phe Glu Val 610 615 620 Val Ala Thr Asn Gly Asp Thr His Leu Gly
Gly Glu Asp Phe Asp Gln 625 630 635 640 Arg Val Met Glu His Phe Ile
Lys Leu Tyr Lys Lys Lys Thr Gly Lys 645 650 655 Asp Val Arg Lys Asp
Asn Arg Ala Val Gln Lys Leu Arg Arg Glu Val 660 665 670 Glu Lys Ala
Lys Arg Ala Leu Ser Ser Gln His Gln Ala Arg Ile Glu 675 680 685 Ile
Glu Ser Phe Tyr Glu Gly Glu Asp Phe Ser Glu Thr Leu Thr Arg 690 695
700 Ala Lys Phe Glu Glu Leu Asn Met Asp Leu Phe Arg Ser Thr Met Lys
705 710 715 720 Pro Val Gln Lys Val Leu Glu Asp Ser Asp Leu Lys Lys
Ser Asp Ile 725 730 735 Asp Glu Ile Val Leu Val Gly Gly Ser Thr Arg
Ile Pro Lys Ile Gln 740 745 750 Gln Leu Val Lys Glu Phe Phe Asn Gly
Lys Glu Pro Ser Arg Gly Ile 755 760 765 Asn Pro Asp Glu Ala Val Ala
Tyr Gly Ala Ala Val Gln Ala Gly Val 770 775 780 Leu Ser Gly Asp Gln
Asp Thr Gly Asp Leu Val Leu Leu Asp Val Cys 785 790 795 800 Pro Leu
Thr Leu Gly Ile Glu Thr Val Gly Gly Val Met Thr Lys Leu 805 810 815
Ile Pro Arg Asn Thr Val Val Pro Thr Lys Lys Ser Gln Ile Phe Ser 820
825 830 Thr Ala Ser Asp Asn Gln Pro Thr Val Thr Ile Lys Val Tyr Glu
Gly 835 840 845 Glu Arg Pro Leu Thr Lys Asp Asn His Leu Leu Gly Thr
Phe Asp Leu 850 855 860 Thr Gly Ile Pro Pro Ala Pro Arg Gly Val Pro
Gln Ile Glu Val Thr 865 870 875 880 Phe Glu Ile Asp Val Asn Gly Ile
Leu Arg Val Thr Ala Glu Asp Lys 885 890 895 Gly Thr Gly Asn Lys Asn
Lys Ile Thr Ile Thr Asn Asp Gln Asn Arg 900 905 910 Leu Thr Pro Glu
Glu Ile Glu Arg Met Val Asn Asp Ala Glu Lys Phe 915 920 925 Ala Glu
Glu Asp Lys Lys Leu Lys Glu Arg Ile Asp Thr Arg Asn Glu 930 935 940
Leu Glu Ser Tyr Ala Tyr Ser Leu Lys Asn Gln Ile Gly Asp Lys Glu 945
950 955 960 Lys Leu Gly Gly Lys Leu Ser Ser Glu Asp Lys Glu Thr Met
Glu Lys 965 970 975 Ala Val Glu Glu Lys Ile Glu Trp Leu Glu Ser His
Gln Asp Ala Asp 980 985 990 Ile Glu Asp Phe Lys Ala Lys Lys Lys Glu
Leu Glu Glu Ile Val Gln 995 1000 1005 Pro Ile Ile Ser Lys Leu Tyr
Gly Ser Ala Gly Pro Pro Pro Thr 1010 1015 1020 Gly Glu Glu Asp Thr
Ala Glu Lys Asp Glu Leu 1025 1030 103345DNAArtificial Sequence3E10
Fv-Hsp90 nucleotide sequence encoding a chimeric protein comprising
a single chain Fv antibody fragment derived from Mus musculus 3E10
hybridoma and Hsp90 from Homo sapiens joined by mouse CH1 linker
and swivel sequence 10atg aga ttt cct tca ctt ttt act gct gtt tta
ttc gca gca tcc tcc 48Met Arg Phe Pro Ser Leu Phe Thr Ala Val Leu
Phe Ala Ala Ser Ser 1 5 10 15 gca tta gct gct cca gtc aac act aca
aca gaa gat gaa acg gca caa 96Ala Leu Ala Ala Pro Val Asn Thr Thr
Thr Glu Asp Glu Thr Ala Gln 20 25 30 att ccg gct gaa gct gtc atc
ggt tac tca gat tta gaa ggg gat ttc 144Ile Pro Ala Glu Ala Val Ile
Gly Tyr Ser Asp Leu Glu Gly Asp Phe 35 40 45 gat gtt gct gtt ttg
cca ttt tcc aac agc aca aat aac ggg tta ttg 192Asp Val Ala Val Leu
Pro Phe Ser Asn Ser Thr Asn Asn Gly Leu Leu 50 55 60 ttt ata aat
act act att gcc agc att gct gct aaa gaa gaa ggg gta 240Phe Ile Asn
Thr Thr Ile Ala Ser Ile Ala Ala Lys Glu Glu Gly Val 65 70 75 80 tct
ctc gag aaa aga gag gct gaa gct gaa ttc cat cac cat cac cat 288Ser
Leu Glu Lys Arg Glu Ala Glu Ala Glu Phe His His His His His 85 90
95 cac gca ggg att cac gac att gtc ctg aca cag tct cct gct tcc tta
336His Ala Gly Ile His Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu
100 105 110 gct gta tct ctg ggg cag agg gcc acc atc tcc tgc agg gcc
agc aaa 384Ala Val Ser Leu Gly Gln Arg Ala Thr Ile Ser Cys Arg Ala
Ser Lys 115 120 125 agt gtc agt aca tct agc tat agt tac atg cac tgg
tac caa cag aaa 432Ser Val Ser Thr Ser Ser Tyr Ser Tyr Met His Trp
Tyr Gln Gln Lys 130 135 140 cca gga cag cca ccc aaa ctc ctc atc aag
tat gca tcc tac cta gaa 480Pro Gly Gln Pro Pro Lys Leu Leu Ile Lys
Tyr Ala Ser Tyr Leu Glu 145 150 155 160 tct ggg gtt cct gcc agg ttc
agt ggc agt ggg tct ggg aca gac ttc 528Ser Gly Val Pro Ala Arg Phe
Ser Gly Ser Gly Ser Gly Thr Asp Phe 165 170 175 acc ctc aac atc cat
cct gtg gag gag gag gat gct gca aca tat tac 576Thr Leu Asn Ile His
Pro Val Glu Glu Glu Asp Ala Ala Thr Tyr Tyr 180 185 190 tgt cag cac
agt agg gag ttt ccg tgg acg ttc ggt gga ggc acc aag 624Cys Gln His
Ser Arg Glu Phe Pro Trp Thr Phe Gly Gly Gly Thr Lys 195 200 205 ctg
gaa atc aaa cgg gct gat gct gca ccc ggg ggt ggc ggt tct ggc 672Leu
Glu Ile Lys Arg Ala Asp Ala Ala Pro Gly Gly Gly Gly Ser Gly 210 215
220 ggt ggc ggt tct gga ggc ggt ggc tct gag gtg cag ctg gtg gag tct
720Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser
225 230 235 240 ggg gga ggc tta gtg aag cct gga ggg tcc cgg aaa ctc
tcc tgt gca 768Gly Gly Gly Leu Val Lys Pro Gly Gly Ser Arg Lys Leu
Ser Cys Ala 245 250 255 gcc tct gga ttc act ttc agt aac tat gga atg
cac tgg gtc cgt cag 816Ala Ser Gly Phe Thr Phe Ser Asn Tyr Gly Met
His Trp Val Arg Gln 260 265 270 gct cca gag aag ggg ctg gag tgg gtt
gca tac att agt agt ggc agt 864Ala Pro Glu Lys Gly Leu Glu Trp Val
Ala Tyr Ile Ser Ser Gly Ser 275 280 285 agt acc atc tac tat gca gac
aca gtg aag ggc cga ttc acc atc tcc 912Ser Thr Ile Tyr Tyr Ala Asp
Thr Val Lys Gly Arg Phe Thr Ile Ser 290 295 300 aga gac aat gcc aag
aac acc ctg ttc ctg caa atg acc agt cta agg 960Arg Asp Asn Ala Lys
Asn Thr Leu Phe Leu Gln Met Thr Ser Leu Arg 305 310 315 320 tct gag
gac aca gcc atg cgg ggg tta cta ctt gac tac tgg ggc caa 1008Ser Glu
Asp Thr Ala Met Arg Gly Leu Leu Leu Asp Tyr Trp Gly Gln 325 330 335
ggc acc act ctc aca gtc tcc tca cta gaa caa aaa ctc atc tca gaa
1056Gly Thr Thr Leu Thr Val Ser Ser Leu Glu Gln Lys Leu Ile Ser Glu
340 345 350 gag gat ctg aat agc gcc gtc gac gcc aaa aca aca gcc cca
tcg gtc 1104Glu Asp Leu Asn Ser Ala Val Asp Ala Lys Thr Thr Ala Pro
Ser Val 355 360 365 tat cca ctg gcc cct gta ctg gag tct tcc gga tcc
atg cct gag gaa 1152Tyr Pro Leu Ala Pro Val Leu Glu Ser Ser Gly Ser
Met Pro Glu Glu 370 375 380 acc cag acc caa gac caa ccg atg gag gag
gag gag gtt gag acg ttc 1200Thr Gln Thr Gln Asp Gln Pro Met Glu Glu
Glu Glu Val Glu Thr Phe 385 390 395 400 gcc ttt cag gca gaa att gcc
cag ttg atg tca ttg atc atc aat act 1248Ala Phe Gln Ala Glu Ile Ala
Gln Leu Met Ser Leu Ile Ile Asn Thr 405 410 415 ttc tac tcg aac aaa
gag atc ttt ctg aga gag ctc att tca aat tca 1296Phe Tyr Ser Asn Lys
Glu Ile Phe Leu Arg Glu Leu Ile Ser Asn Ser 420 425 430 tca gat gca
ttg gac aaa atc cgg tat gaa agc ttg aca gat ccc agt 1344Ser Asp Ala
Leu Asp Lys Ile Arg Tyr Glu Ser Leu Thr Asp Pro Ser 435 440 445 aaa
tta gac tct ggg aaa gag ctg cat att aac ctt ata ccg aac aaa 1392Lys
Leu Asp Ser Gly Lys Glu Leu His Ile Asn Leu Ile Pro Asn Lys 450 455
460 caa gat cga act ctc act att gtg gat act gga att gga atg acc aag
1440Gln Asp Arg Thr Leu Thr Ile Val Asp Thr Gly Ile Gly Met Thr Lys
465 470 475 480 gct gac ttg atc aat aac ctt ggt act atc gcc aag tct
ggg acc aaa 1488Ala Asp Leu Ile Asn Asn Leu Gly Thr Ile Ala Lys Ser
Gly Thr Lys 485 490 495 gcg ttc atg gaa gct ttg cag gct ggt gca gat
atc tct atg att ggc 1536Ala Phe Met Glu Ala Leu Gln Ala Gly Ala Asp
Ile Ser Met Ile Gly 500 505 510 cag ttc ggt gtt ggt ttt tat tct gct
tat ttg gtt gct gag aaa gta 1584Gln Phe Gly Val Gly Phe Tyr Ser Ala
Tyr Leu Val Ala Glu Lys Val 515 520 525 act gtg atc acc aaa cat aac
gat gat gag cag tac gct tgg gag tcc 1632Thr Val Ile Thr Lys His Asn
Asp Asp Glu Gln Tyr Ala Trp Glu Ser 530 535 540 tca gca ggg gga tca
ttc aca gtg agg aca gac aca ggt gaa cct atg 1680Ser Ala Gly Gly Ser
Phe Thr Val Arg Thr Asp Thr Gly Glu Pro Met 545 550 555 560 ggt cgt
gga aca aaa gtt atc cta cac ctg aaa gaa gac caa act gag 1728Gly Arg
Gly Thr Lys Val Ile Leu His Leu Lys Glu Asp Gln Thr Glu 565 570 575
tac ttg gag gaa cga aga ata aag gag att gtg aag aaa cat tct cag
1776Tyr Leu Glu Glu Arg Arg Ile Lys Glu Ile Val Lys Lys His Ser Gln
580 585 590 ttt att gga tat ccc att act ctt ttt gtg gag aag gaa cgt
gat aaa 1824Phe Ile Gly Tyr Pro Ile Thr Leu Phe Val Glu Lys Glu Arg
Asp Lys 595 600 605 gaa gta agc gat gat gag gct gaa gaa aag gaa gac
aaa gaa gaa gaa 1872Glu Val Ser Asp Asp Glu Ala Glu Glu Lys Glu Asp
Lys Glu Glu Glu 610 615 620 aaa gaa aaa gaa gag aaa gag tcg gaa gac
aaa cct gaa att gaa gat 1920Lys Glu Lys Glu Glu Lys Glu Ser Glu Asp
Lys Pro Glu Ile Glu Asp 625 630 635 640 gtt ggt tct gat gag gaa gaa
gaa aag aag gat ggt gac aag aag aag 1968Val Gly Ser Asp Glu Glu Glu
Glu Lys Lys Asp Gly Asp Lys Lys Lys 645 650 655 aag aag aag att aag
gaa aag tac atc gat caa gaa gag ctc aac aaa 2016Lys Lys Lys Ile Lys
Glu Lys Tyr Ile Asp Gln Glu Glu Leu Asn Lys 660 665 670 aca aag ccc
atc tgg acc aga aat ccc gac gat att act aat gag gag 2064Thr Lys Pro
Ile Trp Thr Arg Asn Pro Asp Asp Ile Thr Asn Glu Glu 675 680 685 tac
gga gaa ttc tat aag agc ttg acc aat gac tgg gaa gat cac ttg 2112Tyr
Gly Glu Phe Tyr Lys Ser Leu Thr Asn Asp Trp Glu Asp His Leu 690 695
700 gca gtg aag cat ttt tca gtt gaa gga cag ttg gaa ttc aga gcc ctt
2160Ala Val Lys His Phe Ser Val Glu Gly Gln Leu Glu Phe Arg Ala Leu
705 710 715 720 cta ttt gtc cca cga cgt gct cct ttt gat ctg ttt gaa
aac aga aag 2208Leu Phe Val Pro Arg Arg Ala Pro Phe Asp Leu Phe Glu
Asn Arg Lys 725 730 735 aaa aag aac aac atc aaa ttg tat gta cgc aga
gtt ttc atc atg gat 2256Lys Lys Asn Asn Ile Lys Leu Tyr Val Arg Arg
Val Phe Ile Met Asp 740 745 750 aac tgt gag gag cta atc cct gaa tat
ctg aac ttc att aga ggg gtg 2304Asn Cys Glu Glu Leu Ile Pro Glu Tyr
Leu Asn Phe Ile Arg Gly Val 755 760 765 gta gac tcg gag gat ctc cct
cta aac ata tcc cgt gag atg ttg caa 2352Val Asp Ser Glu Asp Leu Pro
Leu Asn Ile Ser Arg Glu Met Leu Gln 770 775 780 caa agc aaa att ttg
aaa gtt atc agg aag aat ttg gtc aaa aaa tgc 2400Gln Ser Lys Ile Leu
Lys Val Ile Arg Lys Asn Leu Val Lys Lys Cys 785 790 795 800 tta gaa
ctc ttt act gaa ctg gcg gaa gat aaa gag aac tac aag aaa 2448Leu Glu
Leu Phe Thr Glu Leu Ala Glu Asp Lys Glu Asn Tyr Lys Lys 805 810 815
ttc tat gag cag ttc tct aaa aac ata aag ctt gga ata cac gaa
gac 2496Phe Tyr Glu Gln Phe Ser Lys Asn Ile Lys Leu Gly Ile His Glu
Asp 820 825 830 tct caa aat cgg aag aag ctt tca gag ctg tta agg tac
tac aca tct 2544Ser Gln Asn Arg Lys Lys Leu Ser Glu Leu Leu Arg Tyr
Tyr Thr Ser 835 840 845 gcc tct ggt gat gag atg gtt tct ctc aag gac
tac tgc acc aga atg 2592Ala Ser Gly Asp Glu Met Val Ser Leu Lys Asp
Tyr Cys Thr Arg Met 850 855 860 aag gag aac cag aaa cat atc tat tat
atc aca ggt gag acc aag gac 2640Lys Glu Asn Gln Lys His Ile Tyr Tyr
Ile Thr Gly Glu Thr Lys Asp 865 870 875 880 cag gta gct aac tca gcc
ttt gtg gaa cgt ctt cgg aaa cat ggc tta 2688Gln Val Ala Asn Ser Ala
Phe Val Glu Arg Leu Arg Lys His Gly Leu 885 890 895 gaa gtg atc tat
atg att gag ccc att gat gag tac tgt gtc caa cag 2736Glu Val Ile Tyr
Met Ile Glu Pro Ile Asp Glu Tyr Cys Val Gln Gln 900 905 910 ctg aag
gaa ttt gag ggg aag act tta gtg tca gtc acc aaa gaa ggc 2784Leu Lys
Glu Phe Glu Gly Lys Thr Leu Val Ser Val Thr Lys Glu Gly 915 920 925
ctg gaa ctt cca gag gat gaa gaa gag aaa aag aag cag gaa gag aaa
2832Leu Glu Leu Pro Glu Asp Glu Glu Glu Lys Lys Lys Gln Glu Glu Lys
930 935 940 aaa aca aag ttt gag aac ctc tgc aaa atc atg aaa gac ata
ttg gag 2880Lys Thr Lys Phe Glu Asn Leu Cys Lys Ile Met Lys Asp Ile
Leu Glu 945 950 955 960 aaa aaa gtt gaa aag gtg gtt gtg tca aac cga
ttg gtg aca tct cca 2928Lys Lys Val Glu Lys Val Val Val Ser Asn Arg
Leu Val Thr Ser Pro 965 970 975 tgc tgt att gtc aca agc aca tat ggc
tgg aca gca aac atg gag aga 2976Cys Cys Ile Val Thr Ser Thr Tyr Gly
Trp Thr Ala Asn Met Glu Arg 980 985 990 atc atg aaa gct caa gcc cta
aga gac aac tca aca atg ggt tac atg 3024Ile Met Lys Ala Gln Ala Leu
Arg Asp Asn Ser Thr Met Gly Tyr Met 995 1000 1005 gca gca aag aaa
cac ctg gag ata aac cct gac cat tcc att att 3069Ala Ala Lys Lys His
Leu Glu Ile Asn Pro Asp His Ser Ile Ile 1010 1015 1020 gag acc tta
agg caa aag gca gag gct gat aag aac gac aag tct 3114Glu Thr Leu Arg
Gln Lys Ala Glu Ala Asp Lys Asn Asp Lys Ser 1025 1030 1035 gtg aag
gat ctg gtc atc ttg ctt tat gaa act gcg ctc ctg tct 3159Val Lys Asp
Leu Val Ile Leu Leu Tyr Glu Thr Ala Leu Leu Ser 1040 1045 1050 tct
ggc ttc agt ctg gaa gat ccc cag aca cat gct aac agg atc 3204Ser Gly
Phe Ser Leu Glu Asp Pro Gln Thr His Ala Asn Arg Ile 1055 1060 1065
tac agg atg atc aaa ctt ggt ctg ggt att gat gaa gat gac cct 3249Tyr
Arg Met Ile Lys Leu Gly Leu Gly Ile Asp Glu Asp Asp Pro 1070 1075
1080 act gct gat gat acc agt gct gct gta act gaa gaa atg cca ccc
3294Thr Ala Asp Asp Thr Ser Ala Ala Val Thr Glu Glu Met Pro Pro
1085 1090 1095 ctt gaa gga gat gac gac aca tca cgc atg gaa gaa gta
gac taa 3339Leu Glu Gly Asp Asp Asp Thr Ser Arg Met Glu Glu Val Asp
1100 1105 1110 tctaga 3345111112PRTArtificial SequenceSynthetic
Construct 11Met Arg Phe Pro Ser Leu Phe Thr Ala Val Leu Phe Ala Ala
Ser Ser 1 5 10 15 Ala Leu Ala Ala Pro Val Asn Thr Thr Thr Glu Asp
Glu Thr Ala Gln 20 25 30 Ile Pro Ala Glu Ala Val Ile Gly Tyr Ser
Asp Leu Glu Gly Asp Phe 35 40 45 Asp Val Ala Val Leu Pro Phe Ser
Asn Ser Thr Asn Asn Gly Leu Leu 50 55 60 Phe Ile Asn Thr Thr Ile
Ala Ser Ile Ala Ala Lys Glu Glu Gly Val 65 70 75 80 Ser Leu Glu Lys
Arg Glu Ala Glu Ala Glu Phe His His His His His 85 90 95 His Ala
Gly Ile His Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu 100 105 110
Ala Val Ser Leu Gly Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Lys 115
120 125 Ser Val Ser Thr Ser Ser Tyr Ser Tyr Met His Trp Tyr Gln Gln
Lys 130 135 140 Pro Gly Gln Pro Pro Lys Leu Leu Ile Lys Tyr Ala Ser
Tyr Leu Glu 145 150 155 160 Ser Gly Val Pro Ala Arg Phe Ser Gly Ser
Gly Ser Gly Thr Asp Phe 165 170 175 Thr Leu Asn Ile His Pro Val Glu
Glu Glu Asp Ala Ala Thr Tyr Tyr 180 185 190 Cys Gln His Ser Arg Glu
Phe Pro Trp Thr Phe Gly Gly Gly Thr Lys 195 200 205 Leu Glu Ile Lys
Arg Ala Asp Ala Ala Pro Gly Gly Gly Gly Ser Gly 210 215 220 Gly Gly
Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser 225 230 235
240 Gly Gly Gly Leu Val Lys Pro Gly Gly Ser Arg Lys Leu Ser Cys Ala
245 250 255 Ala Ser Gly Phe Thr Phe Ser Asn Tyr Gly Met His Trp Val
Arg Gln 260 265 270 Ala Pro Glu Lys Gly Leu Glu Trp Val Ala Tyr Ile
Ser Ser Gly Ser 275 280 285 Ser Thr Ile Tyr Tyr Ala Asp Thr Val Lys
Gly Arg Phe Thr Ile Ser 290 295 300 Arg Asp Asn Ala Lys Asn Thr Leu
Phe Leu Gln Met Thr Ser Leu Arg 305 310 315 320 Ser Glu Asp Thr Ala
Met Arg Gly Leu Leu Leu Asp Tyr Trp Gly Gln 325 330 335 Gly Thr Thr
Leu Thr Val Ser Ser Leu Glu Gln Lys Leu Ile Ser Glu 340 345 350 Glu
Asp Leu Asn Ser Ala Val Asp Ala Lys Thr Thr Ala Pro Ser Val 355 360
365 Tyr Pro Leu Ala Pro Val Leu Glu Ser Ser Gly Ser Met Pro Glu Glu
370 375 380 Thr Gln Thr Gln Asp Gln Pro Met Glu Glu Glu Glu Val Glu
Thr Phe 385 390 395 400 Ala Phe Gln Ala Glu Ile Ala Gln Leu Met Ser
Leu Ile Ile Asn Thr 405 410 415 Phe Tyr Ser Asn Lys Glu Ile Phe Leu
Arg Glu Leu Ile Ser Asn Ser 420 425 430 Ser Asp Ala Leu Asp Lys Ile
Arg Tyr Glu Ser Leu Thr Asp Pro Ser 435 440 445 Lys Leu Asp Ser Gly
Lys Glu Leu His Ile Asn Leu Ile Pro Asn Lys 450 455 460 Gln Asp Arg
Thr Leu Thr Ile Val Asp Thr Gly Ile Gly Met Thr Lys 465 470 475 480
Ala Asp Leu Ile Asn Asn Leu Gly Thr Ile Ala Lys Ser Gly Thr Lys 485
490 495 Ala Phe Met Glu Ala Leu Gln Ala Gly Ala Asp Ile Ser Met Ile
Gly 500 505 510 Gln Phe Gly Val Gly Phe Tyr Ser Ala Tyr Leu Val Ala
Glu Lys Val 515 520 525 Thr Val Ile Thr Lys His Asn Asp Asp Glu Gln
Tyr Ala Trp Glu Ser 530 535 540 Ser Ala Gly Gly Ser Phe Thr Val Arg
Thr Asp Thr Gly Glu Pro Met 545 550 555 560 Gly Arg Gly Thr Lys Val
Ile Leu His Leu Lys Glu Asp Gln Thr Glu 565 570 575 Tyr Leu Glu Glu
Arg Arg Ile Lys Glu Ile Val Lys Lys His Ser Gln 580 585 590 Phe Ile
Gly Tyr Pro Ile Thr Leu Phe Val Glu Lys Glu Arg Asp Lys 595 600 605
Glu Val Ser Asp Asp Glu Ala Glu Glu Lys Glu Asp Lys Glu Glu Glu 610
615 620 Lys Glu Lys Glu Glu Lys Glu Ser Glu Asp Lys Pro Glu Ile Glu
Asp 625 630 635 640 Val Gly Ser Asp Glu Glu Glu Glu Lys Lys Asp Gly
Asp Lys Lys Lys 645 650 655 Lys Lys Lys Ile Lys Glu Lys Tyr Ile Asp
Gln Glu Glu Leu Asn Lys 660 665 670 Thr Lys Pro Ile Trp Thr Arg Asn
Pro Asp Asp Ile Thr Asn Glu Glu 675 680 685 Tyr Gly Glu Phe Tyr Lys
Ser Leu Thr Asn Asp Trp Glu Asp His Leu 690 695 700 Ala Val Lys His
Phe Ser Val Glu Gly Gln Leu Glu Phe Arg Ala Leu 705 710 715 720 Leu
Phe Val Pro Arg Arg Ala Pro Phe Asp Leu Phe Glu Asn Arg Lys 725 730
735 Lys Lys Asn Asn Ile Lys Leu Tyr Val Arg Arg Val Phe Ile Met Asp
740 745 750 Asn Cys Glu Glu Leu Ile Pro Glu Tyr Leu Asn Phe Ile Arg
Gly Val 755 760 765 Val Asp Ser Glu Asp Leu Pro Leu Asn Ile Ser Arg
Glu Met Leu Gln 770 775 780 Gln Ser Lys Ile Leu Lys Val Ile Arg Lys
Asn Leu Val Lys Lys Cys 785 790 795 800 Leu Glu Leu Phe Thr Glu Leu
Ala Glu Asp Lys Glu Asn Tyr Lys Lys 805 810 815 Phe Tyr Glu Gln Phe
Ser Lys Asn Ile Lys Leu Gly Ile His Glu Asp 820 825 830 Ser Gln Asn
Arg Lys Lys Leu Ser Glu Leu Leu Arg Tyr Tyr Thr Ser 835 840 845 Ala
Ser Gly Asp Glu Met Val Ser Leu Lys Asp Tyr Cys Thr Arg Met 850 855
860 Lys Glu Asn Gln Lys His Ile Tyr Tyr Ile Thr Gly Glu Thr Lys Asp
865 870 875 880 Gln Val Ala Asn Ser Ala Phe Val Glu Arg Leu Arg Lys
His Gly Leu 885 890 895 Glu Val Ile Tyr Met Ile Glu Pro Ile Asp Glu
Tyr Cys Val Gln Gln 900 905 910 Leu Lys Glu Phe Glu Gly Lys Thr Leu
Val Ser Val Thr Lys Glu Gly 915 920 925 Leu Glu Leu Pro Glu Asp Glu
Glu Glu Lys Lys Lys Gln Glu Glu Lys 930 935 940 Lys Thr Lys Phe Glu
Asn Leu Cys Lys Ile Met Lys Asp Ile Leu Glu 945 950 955 960 Lys Lys
Val Glu Lys Val Val Val Ser Asn Arg Leu Val Thr Ser Pro 965 970 975
Cys Cys Ile Val Thr Ser Thr Tyr Gly Trp Thr Ala Asn Met Glu Arg 980
985 990 Ile Met Lys Ala Gln Ala Leu Arg Asp Asn Ser Thr Met Gly Tyr
Met 995 1000 1005 Ala Ala Lys Lys His Leu Glu Ile Asn Pro Asp His
Ser Ile Ile 1010 1015 1020 Glu Thr Leu Arg Gln Lys Ala Glu Ala Asp
Lys Asn Asp Lys Ser 1025 1030 1035 Val Lys Asp Leu Val Ile Leu Leu
Tyr Glu Thr Ala Leu Leu Ser 1040 1045 1050 Ser Gly Phe Ser Leu Glu
Asp Pro Gln Thr His Ala Asn Arg Ile 1055 1060 1065 Tyr Arg Met Ile
Lys Leu Gly Leu Gly Ile Asp Glu Asp Asp Pro 1070 1075 1080 Thr Ala
Asp Asp Thr Ser Ala Ala Val Thr Glu Glu Met Pro Pro 1085 1090 1095
Leu Glu Gly Asp Asp Asp Thr Ser Arg Met Glu Glu Val Asp 1100 1105
1110 121112PRTArtificial Sequence3E10 Fv-Hsp90 amino acid sequence
for a chimeric protein comprising a single chain Fv antibody
fragment derived from Mus musculus 3E10 hybridoma and Hsp90 from
Homo sapiens joined by mouse CH1 linker and swivel sequence 12Met
Arg Phe Pro Ser Leu Phe Thr Ala Val Leu Phe Ala Ala Ser Ser 1 5 10
15 Ala Leu Ala Ala Pro Val Asn Thr Thr Thr Glu Asp Glu Thr Ala Gln
20 25 30 Ile Pro Ala Glu Ala Val Ile Gly Tyr Ser Asp Leu Glu Gly
Asp Phe 35 40 45 Asp Val Ala Val Leu Pro Phe Ser Asn Ser Thr Asn
Asn Gly Leu Leu 50 55 60 Phe Ile Asn Thr Thr Ile Ala Ser Ile Ala
Ala Lys Glu Glu Gly Val 65 70 75 80 Ser Leu Glu Lys Arg Glu Ala Glu
Ala Glu Phe His His His His His 85 90 95 His Ala Gly Ile His Asp
Ile Val Leu Thr Gln Ser Pro Ala Ser Leu 100 105 110 Ala Val Ser Leu
Gly Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Lys 115 120 125 Ser Val
Ser Thr Ser Ser Tyr Ser Tyr Met His Trp Tyr Gln Gln Lys 130 135 140
Pro Gly Gln Pro Pro Lys Leu Leu Ile Lys Tyr Ala Ser Tyr Leu Glu 145
150 155 160 Ser Gly Val Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr
Asp Phe 165 170 175 Thr Leu Asn Ile His Pro Val Glu Glu Glu Asp Ala
Ala Thr Tyr Tyr 180 185 190 Cys Gln His Ser Arg Glu Phe Pro Trp Thr
Phe Gly Gly Gly Thr Lys 195 200 205 Leu Glu Ile Lys Arg Ala Asp Ala
Ala Pro Gly Gly Gly Gly Ser Gly 210 215 220 Gly Gly Gly Ser Gly Gly
Gly Gly Ser Glu Val Gln Leu Val Glu Ser 225 230 235 240 Gly Gly Gly
Leu Val Lys Pro Gly Gly Ser Arg Lys Leu Ser Cys Ala 245 250 255 Ala
Ser Gly Phe Thr Phe Ser Asn Tyr Gly Met His Trp Val Arg Gln 260 265
270 Ala Pro Glu Lys Gly Leu Glu Trp Val Ala Tyr Ile Ser Ser Gly Ser
275 280 285 Ser Thr Ile Tyr Tyr Ala Asp Thr Val Lys Gly Arg Phe Thr
Ile Ser 290 295 300 Arg Asp Asn Ala Lys Asn Thr Leu Phe Leu Gln Met
Thr Ser Leu Arg 305 310 315 320 Ser Glu Asp Thr Ala Met Arg Gly Leu
Leu Leu Asp Tyr Trp Gly Gln 325 330 335 Gly Thr Thr Leu Thr Val Ser
Ser Leu Glu Gln Lys Leu Ile Ser Glu 340 345 350 Glu Asp Leu Asn Ser
Ala Val Asp Ala Lys Thr Thr Ala Pro Ser Val 355 360 365 Tyr Pro Leu
Ala Pro Val Leu Glu Ser Ser Gly Ser Met Pro Glu Glu 370 375 380 Thr
Gln Thr Gln Asp Gln Pro Met Glu Glu Glu Glu Val Glu Thr Phe 385 390
395 400 Ala Phe Gln Ala Glu Ile Ala Gln Leu Met Ser Leu Ile Ile Asn
Thr 405 410 415 Phe Tyr Ser Asn Lys Glu Ile Phe Leu Arg Glu Leu Ile
Ser Asn Ser 420 425 430 Ser Asp Ala Leu Asp Lys Ile Arg Tyr Glu Ser
Leu Thr Asp Pro Ser 435 440 445 Lys Leu Asp Ser Gly Lys Glu Leu His
Ile Asn Leu Ile Pro Asn Lys 450 455 460 Gln Asp Arg Thr Leu Thr Ile
Val Asp Thr Gly Ile Gly Met Thr Lys 465 470 475 480 Ala Asp Leu Ile
Asn Asn Leu Gly Thr Ile Ala Lys Ser Gly Thr Lys 485 490 495 Ala Phe
Met Glu Ala Leu Gln Ala Gly Ala Asp Ile Ser Met Ile Gly 500 505 510
Gln Phe Gly Val Gly Phe Tyr Ser Ala Tyr Leu Val Ala Glu Lys Val 515
520 525 Thr Val Ile Thr Lys His Asn Asp Asp Glu Gln Tyr Ala Trp Glu
Ser 530 535 540 Ser Ala Gly Gly Ser Phe Thr Val Arg Thr Asp Thr Gly
Glu Pro Met 545 550 555 560 Gly Arg Gly Thr Lys Val Ile Leu His Leu
Lys Glu Asp Gln Thr Glu 565 570 575 Tyr Leu Glu Glu Arg Arg Ile Lys
Glu Ile Val Lys Lys His Ser Gln 580 585 590 Phe Ile Gly Tyr Pro Ile
Thr Leu Phe Val Glu Lys Glu Arg Asp Lys 595 600 605 Glu Val Ser Asp
Asp Glu Ala Glu Glu Lys Glu Asp Lys Glu Glu Glu 610
615 620 Lys Glu Lys Glu Glu Lys Glu Ser Glu Asp Lys Pro Glu Ile Glu
Asp 625 630 635 640 Val Gly Ser Asp Glu Glu Glu Glu Lys Lys Asp Gly
Asp Lys Lys Lys 645 650 655 Lys Lys Lys Ile Lys Glu Lys Tyr Ile Asp
Gln Glu Glu Leu Asn Lys 660 665 670 Thr Lys Pro Ile Trp Thr Arg Asn
Pro Asp Asp Ile Thr Asn Glu Glu 675 680 685 Tyr Gly Glu Phe Tyr Lys
Ser Leu Thr Asn Asp Trp Glu Asp His Leu 690 695 700 Ala Val Lys His
Phe Ser Val Glu Gly Gln Leu Glu Phe Arg Ala Leu 705 710 715 720 Leu
Phe Val Pro Arg Arg Ala Pro Phe Asp Leu Phe Glu Asn Arg Lys 725 730
735 Lys Lys Asn Asn Ile Lys Leu Tyr Val Arg Arg Val Phe Ile Met Asp
740 745 750 Asn Cys Glu Glu Leu Ile Pro Glu Tyr Leu Asn Phe Ile Arg
Gly Val 755 760 765 Val Asp Ser Glu Asp Leu Pro Leu Asn Ile Ser Arg
Glu Met Leu Gln 770 775 780 Gln Ser Lys Ile Leu Lys Val Ile Arg Lys
Asn Leu Val Lys Lys Cys 785 790 795 800 Leu Glu Leu Phe Thr Glu Leu
Ala Glu Asp Lys Glu Asn Tyr Lys Lys 805 810 815 Phe Tyr Glu Gln Phe
Ser Lys Asn Ile Lys Leu Gly Ile His Glu Asp 820 825 830 Ser Gln Asn
Arg Lys Lys Leu Ser Glu Leu Leu Arg Tyr Tyr Thr Ser 835 840 845 Ala
Ser Gly Asp Glu Met Val Ser Leu Lys Asp Tyr Cys Thr Arg Met 850 855
860 Lys Glu Asn Gln Lys His Ile Tyr Tyr Ile Thr Gly Glu Thr Lys Asp
865 870 875 880 Gln Val Ala Asn Ser Ala Phe Val Glu Arg Leu Arg Lys
His Gly Leu 885 890 895 Glu Val Ile Tyr Met Ile Glu Pro Ile Asp Glu
Tyr Cys Val Gln Gln 900 905 910 Leu Lys Glu Phe Glu Gly Lys Thr Leu
Val Ser Val Thr Lys Glu Gly 915 920 925 Leu Glu Leu Pro Glu Asp Glu
Glu Glu Lys Lys Lys Gln Glu Glu Lys 930 935 940 Lys Thr Lys Phe Glu
Asn Leu Cys Lys Ile Met Lys Asp Ile Leu Glu 945 950 955 960 Lys Lys
Val Glu Lys Val Val Val Ser Asn Arg Leu Val Thr Ser Pro 965 970 975
Cys Cys Ile Val Thr Ser Thr Tyr Gly Trp Thr Ala Asn Met Glu Arg 980
985 990 Ile Met Lys Ala Gln Ala Leu Arg Asp Asn Ser Thr Met Gly Tyr
Met 995 1000 1005 Ala Ala Lys Lys His Leu Glu Ile Asn Pro Asp His
Ser Ile Ile 1010 1015 1020 Glu Thr Leu Arg Gln Lys Ala Glu Ala Asp
Lys Asn Asp Lys Ser 1025 1030 1035 Val Lys Asp Leu Val Ile Leu Leu
Tyr Glu Thr Ala Leu Leu Ser 1040 1045 1050 Ser Gly Phe Ser Leu Glu
Asp Pro Gln Thr His Ala Asn Arg Ile 1055 1060 1065 Tyr Arg Met Ile
Lys Leu Gly Leu Gly Ile Asp Glu Asp Asp Pro 1070 1075 1080 Thr Ala
Asp Asp Thr Ser Ala Ala Val Thr Glu Glu Met Pro Pro 1085 1090 1095
Leu Glu Gly Asp Asp Asp Thr Ser Arg Met Glu Glu Val Asp 1100 1105
1110 1312DNAMus musculusCDS(1)..(12)anti-DNA monoclonal antibody
3E10 kappa light (Vk) chain complementarity determining region 1
(CDR1) coding sequence 13agt tac atg cac 12Ser Tyr Met His 1
144PRTMus musculus 14Ser Tyr Met His 1 1518DNAMus
musculusCDS(1)..(18)anti-DNA monoclonal antibody 3E10 kappa light
(Vk) chain complementarity determining region 2 (CDR2) coding
sequence 15gca tcc tac cta gaa tct 18Ala Ser Tyr Leu Glu Ser 1 5
166PRTMus musculus 16Ala Ser Tyr Leu Glu Ser 1 5 1727DNAMus
musculusCDS(1)..(27)anti-DNA monoclonal antibody 3E10 kappa light
(Vk) chain complementarity determining region 3 (CDR3) coding
sequence 17cag cac agt agg gag ttt ccg tgg acg 27Gln His Ser Arg
Glu Phe Pro Trp Thr 1 5 189PRTMus musculus 18Gln His Ser Arg Glu
Phe Pro Trp Thr 1 5 1915DNAMus musculusCDS(1)..(15)anti-DNA
monoclonal antibody 3E10 variable heavy (VH) chain complementarity
determining region 1 (CDR1) coding sequence with D31N mutation at
the first amino acid position of CDR1 for enhanced cell penetration
19aac tat gga atg cac 15Asn Tyr Gly Met His 1 5 205PRTMus musculus
20Asn Tyr Gly Met His 1 5 2151DNAMus musculusCDS(1)..(51)anti-DNA
monoclonal antibody 3E10 variable heavy (VH) chain complementarity
determining region 2 (CDR2) coding sequence 21tac att agt agt ggc
agt agt acc atc tac tat gca gac aca gtg aag 48Tyr Ile Ser Ser Gly
Ser Ser Thr Ile Tyr Tyr Ala Asp Thr Val Lys 1 5 10 15 ggc 51Gly
2217PRTMus musculus 22Tyr Ile Ser Ser Gly Ser Ser Thr Ile Tyr Tyr
Ala Asp Thr Val Lys 1 5 10 15 Gly 2321DNAMus
musculusCDS(1)..(21)anti-DNA monoclonal antibody 3E10 variable
heavy (VH) chain complementarity determining region 3 (CDR3) coding
sequence 23cgg ggg tta cta ctt gac tac 21Arg Gly Leu Leu Leu Asp
Tyr 1 5 247PRTMus musculus 24Arg Gly Leu Leu Leu Asp Tyr 1 5
251761DNAArtificial Sequence3E10 Fv-Hsp27 nucleotide sequence
encoding a chimeric protein comprising a single chain Fv antibody
fragment derived from Mus musculus 3E10 hybridoma and Hsp27 from
Homo sapiens joined by human CH1 linker and swivel sequence 25atg
aga ttt cct tca ctt ttt act gct gtt tta ttc gca gca tcc tcc 48Met
Arg Phe Pro Ser Leu Phe Thr Ala Val Leu Phe Ala Ala Ser Ser 1 5 10
15 gca tta gct gct cca gtc aac act aca aca gaa gat gaa acg gca caa
96Ala Leu Ala Ala Pro Val Asn Thr Thr Thr Glu Asp Glu Thr Ala Gln
20 25 30 att ccg gct gaa gct gtc atc ggt tac tca gat tta gaa ggg
gat ttc 144Ile Pro Ala Glu Ala Val Ile Gly Tyr Ser Asp Leu Glu Gly
Asp Phe 35 40 45 gat gtt gct gtt ttg cca ttt tcc aac agc aca aat
aac ggg tta ttg 192Asp Val Ala Val Leu Pro Phe Ser Asn Ser Thr Asn
Asn Gly Leu Leu 50 55 60 ttt ata aat act act att gcc agc att gct
gct aaa gaa gaa ggg gta 240Phe Ile Asn Thr Thr Ile Ala Ser Ile Ala
Ala Lys Glu Glu Gly Val 65 70 75 80 tct ctc gag aaa aga gag gct gaa
gct gaa ttc cat cac cat cac cat 288Ser Leu Glu Lys Arg Glu Ala Glu
Ala Glu Phe His His His His His 85 90 95 cac gca ggg att cac gac
att gtc ctg aca cag tct cct gct tcc tta 336His Ala Gly Ile His Asp
Ile Val Leu Thr Gln Ser Pro Ala Ser Leu 100 105 110 gct gta tct ctg
ggg cag agg gcc acc atc tcc tgc agg gcc agc aaa 384Ala Val Ser Leu
Gly Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Lys 115 120 125 agt gtc
agt aca tct agc tat agt tac atg cac tgg tac caa cag aaa 432Ser Val
Ser Thr Ser Ser Tyr Ser Tyr Met His Trp Tyr Gln Gln Lys 130 135 140
cca gga cag cca ccc aaa ctc ctc atc aag tat gca tcc tac cta gaa
480Pro Gly Gln Pro Pro Lys Leu Leu Ile Lys Tyr Ala Ser Tyr Leu Glu
145 150 155 160 tct ggg gtt cct gcc agg ttc agt ggc agt ggg tct ggg
aca gac ttc 528Ser Gly Val Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly
Thr Asp Phe 165 170 175 acc ctc aac atc cat cct gtg gag gag gag gat
gct gca aca tat tac 576Thr Leu Asn Ile His Pro Val Glu Glu Glu Asp
Ala Ala Thr Tyr Tyr 180 185 190 tgt cag cac agt agg gag ttt ccg tgg
acg ttc ggt gga ggc acc aag 624Cys Gln His Ser Arg Glu Phe Pro Trp
Thr Phe Gly Gly Gly Thr Lys 195 200 205 ctg gaa atc aaa cgg gct gat
gct gca ccc ggg ggt ggc ggt tct ggc 672Leu Glu Ile Lys Arg Ala Asp
Ala Ala Pro Gly Gly Gly Gly Ser Gly 210 215 220 ggt ggc ggt tct gga
ggc ggt ggc tct gag gtg cag ctg gtg gag tct 720Gly Gly Gly Ser Gly
Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser 225 230 235 240 ggg gga
ggc tta gtg aag cct gga ggg tcc cgg aaa ctc tcc tgt gca 768Gly Gly
Gly Leu Val Lys Pro Gly Gly Ser Arg Lys Leu Ser Cys Ala 245 250 255
gcc tct gga ttc act ttc agt aac tat gga atg cac tgg gtc cgt cag
816Ala Ser Gly Phe Thr Phe Ser Asn Tyr Gly Met His Trp Val Arg Gln
260 265 270 gct cca gag aag ggg ctg gag tgg gtt gca tac att agt agt
ggc agt 864Ala Pro Glu Lys Gly Leu Glu Trp Val Ala Tyr Ile Ser Ser
Gly Ser 275 280 285 agt acc atc tac tat gca gac aca gtg aag ggc cga
ttc acc atc tcc 912Ser Thr Ile Tyr Tyr Ala Asp Thr Val Lys Gly Arg
Phe Thr Ile Ser 290 295 300 aga gac aat gcc aag aac acc ctg ttc ctg
caa atg acc agt cta agg 960Arg Asp Asn Ala Lys Asn Thr Leu Phe Leu
Gln Met Thr Ser Leu Arg 305 310 315 320 tct gag gac aca gcc atg cgg
ggg tta cta ctt gac tac tgg ggc caa 1008Ser Glu Asp Thr Ala Met Arg
Gly Leu Leu Leu Asp Tyr Trp Gly Gln 325 330 335 ggc acc act ctc aca
gtc tcc tca cta gaa caa aaa ctc atc tca gaa 1056Gly Thr Thr Leu Thr
Val Ser Ser Leu Glu Gln Lys Leu Ile Ser Glu 340 345 350 gag gat ctg
aat agc gcc gtc gac gct tcc acc aag ggc cca tcc gtc 1104Glu Asp Leu
Asn Ser Ala Val Asp Ala Ser Thr Lys Gly Pro Ser Val 355 360 365 ttc
ccc ctg gcg ccc ctg gag tct tcc gga tcc atg acc gag cgc cgc 1152Phe
Pro Leu Ala Pro Leu Glu Ser Ser Gly Ser Met Thr Glu Arg Arg 370 375
380 gtc ccc ttc tcg ctc ctg cgg ggc ccc agc tgg gac ccc ttc cgc gac
1200Val Pro Phe Ser Leu Leu Arg Gly Pro Ser Trp Asp Pro Phe Arg Asp
385 390 395 400 tgg tac ccg cat agc cgc ctc ttc gac cag gcc ttc ggg
ctg ccc cgg 1248Trp Tyr Pro His Ser Arg Leu Phe Asp Gln Ala Phe Gly
Leu Pro Arg 405 410 415 ctg ccg gag gag tgg tcg cag tgg tta ggc ggc
agc agc tgg cca ggc 1296Leu Pro Glu Glu Trp Ser Gln Trp Leu Gly Gly
Ser Ser Trp Pro Gly 420 425 430 tac gtg cgc ccc ctg ccc ccc gcc gcc
atc gag agc ccc gca gtg gcc 1344Tyr Val Arg Pro Leu Pro Pro Ala Ala
Ile Glu Ser Pro Ala Val Ala 435 440 445 gcg ccc gcc tac agc cgc gcg
ctc agc cgg caa ctc agc agc ggg gtc 1392Ala Pro Ala Tyr Ser Arg Ala
Leu Ser Arg Gln Leu Ser Ser Gly Val 450 455 460 tcg gag atc cgg cac
act gcg gac cgc tgg cgc gtg tcc ctg gat gtc 1440Ser Glu Ile Arg His
Thr Ala Asp Arg Trp Arg Val Ser Leu Asp Val 465 470 475 480 aac cac
ttc gcc ccg gac gag ctg acg gtc aag acc aag gat ggc gtg 1488Asn His
Phe Ala Pro Asp Glu Leu Thr Val Lys Thr Lys Asp Gly Val 485 490 495
gtg gag atc acc ggc aag cac gag gag cgg cag gac gag cat ggc tac
1536Val Glu Ile Thr Gly Lys His Glu Glu Arg Gln Asp Glu His Gly Tyr
500 505 510 atc tcc cgg tgc ttc acg cgg aaa tac acg ctg ccc ccc ggt
gtg gac 1584Ile Ser Arg Cys Phe Thr Arg Lys Tyr Thr Leu Pro Pro Gly
Val Asp 515 520 525 ccc acc caa gtt tcc tcc tcc ctg tcc cct gag ggc
aca ctg acc gtg 1632Pro Thr Gln Val Ser Ser Ser Leu Ser Pro Glu Gly
Thr Leu Thr Val 530 535 540 gag gcc ccc atg ccc aag cta gcc acg cag
tcc aac gag atc acc atc 1680Glu Ala Pro Met Pro Lys Leu Ala Thr Gln
Ser Asn Glu Ile Thr Ile 545 550 555 560 cca gtc acc ttc gag tcg cgg
gcc cag ctt ggg ggc cca gaa gct gca 1728Pro Val Thr Phe Glu Ser Arg
Ala Gln Leu Gly Gly Pro Glu Ala Ala 565 570 575 aaa tcc gat gag act
gcc gcc aag taa tctaga 1761Lys Ser Asp Glu Thr Ala Ala Lys 580
26584PRTArtificial SequenceSynthetic Construct 26Met Arg Phe Pro
Ser Leu Phe Thr Ala Val Leu Phe Ala Ala Ser Ser 1 5 10 15 Ala Leu
Ala Ala Pro Val Asn Thr Thr Thr Glu Asp Glu Thr Ala Gln 20 25 30
Ile Pro Ala Glu Ala Val Ile Gly Tyr Ser Asp Leu Glu Gly Asp Phe 35
40 45 Asp Val Ala Val Leu Pro Phe Ser Asn Ser Thr Asn Asn Gly Leu
Leu 50 55 60 Phe Ile Asn Thr Thr Ile Ala Ser Ile Ala Ala Lys Glu
Glu Gly Val 65 70 75 80 Ser Leu Glu Lys Arg Glu Ala Glu Ala Glu Phe
His His His His His 85 90 95 His Ala Gly Ile His Asp Ile Val Leu
Thr Gln Ser Pro Ala Ser Leu 100 105 110 Ala Val Ser Leu Gly Gln Arg
Ala Thr Ile Ser Cys Arg Ala Ser Lys 115 120 125 Ser Val Ser Thr Ser
Ser Tyr Ser Tyr Met His Trp Tyr Gln Gln Lys 130 135 140 Pro Gly Gln
Pro Pro Lys Leu Leu Ile Lys Tyr Ala Ser Tyr Leu Glu 145 150 155 160
Ser Gly Val Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe 165
170 175 Thr Leu Asn Ile His Pro Val Glu Glu Glu Asp Ala Ala Thr Tyr
Tyr 180 185 190 Cys Gln His Ser Arg Glu Phe Pro Trp Thr Phe Gly Gly
Gly Thr Lys 195 200 205 Leu Glu Ile Lys Arg Ala Asp Ala Ala Pro Gly
Gly Gly Gly Ser Gly 210 215 220 Gly Gly Gly Ser Gly Gly Gly Gly Ser
Glu Val Gln Leu Val Glu Ser 225 230 235 240 Gly Gly Gly Leu Val Lys
Pro Gly Gly Ser Arg Lys Leu Ser Cys Ala 245 250 255 Ala Ser Gly Phe
Thr Phe Ser Asn Tyr Gly Met His Trp Val Arg Gln 260 265 270 Ala Pro
Glu Lys Gly Leu Glu Trp Val Ala Tyr Ile Ser Ser Gly Ser 275 280 285
Ser Thr Ile Tyr Tyr Ala Asp Thr Val Lys Gly Arg Phe Thr Ile Ser 290
295 300 Arg Asp Asn Ala Lys Asn Thr Leu Phe Leu Gln Met Thr Ser Leu
Arg 305 310 315 320 Ser Glu Asp Thr Ala Met Arg Gly Leu Leu Leu Asp
Tyr Trp Gly Gln 325 330 335 Gly Thr Thr Leu Thr Val Ser Ser Leu Glu
Gln Lys Leu Ile Ser Glu 340 345 350 Glu Asp Leu Asn Ser Ala Val Asp
Ala Ser Thr Lys Gly Pro Ser Val 355
360 365 Phe Pro Leu Ala Pro Leu Glu Ser Ser Gly Ser Met Thr Glu Arg
Arg 370 375 380 Val Pro Phe Ser Leu Leu Arg Gly Pro Ser Trp Asp Pro
Phe Arg Asp 385 390 395 400 Trp Tyr Pro His Ser Arg Leu Phe Asp Gln
Ala Phe Gly Leu Pro Arg 405 410 415 Leu Pro Glu Glu Trp Ser Gln Trp
Leu Gly Gly Ser Ser Trp Pro Gly 420 425 430 Tyr Val Arg Pro Leu Pro
Pro Ala Ala Ile Glu Ser Pro Ala Val Ala 435 440 445 Ala Pro Ala Tyr
Ser Arg Ala Leu Ser Arg Gln Leu Ser Ser Gly Val 450 455 460 Ser Glu
Ile Arg His Thr Ala Asp Arg Trp Arg Val Ser Leu Asp Val 465 470 475
480 Asn His Phe Ala Pro Asp Glu Leu Thr Val Lys Thr Lys Asp Gly Val
485 490 495 Val Glu Ile Thr Gly Lys His Glu Glu Arg Gln Asp Glu His
Gly Tyr 500 505 510 Ile Ser Arg Cys Phe Thr Arg Lys Tyr Thr Leu Pro
Pro Gly Val Asp 515 520 525 Pro Thr Gln Val Ser Ser Ser Leu Ser Pro
Glu Gly Thr Leu Thr Val 530 535 540 Glu Ala Pro Met Pro Lys Leu Ala
Thr Gln Ser Asn Glu Ile Thr Ile 545 550 555 560 Pro Val Thr Phe Glu
Ser Arg Ala Gln Leu Gly Gly Pro Glu Ala Ala 565 570 575 Lys Ser Asp
Glu Thr Ala Ala Lys 580 27584PRTArtificial Sequence3E10 Fv-Hsp27
amino acid sequence for a chimeric protein comprising a single
chain Fv antibody fragment derived from Mus musculus 3E10 hybridoma
and Hsp27 from Homo sapiens joined by human CH1 linker and swivel
sequence 27Met Arg Phe Pro Ser Leu Phe Thr Ala Val Leu Phe Ala Ala
Ser Ser 1 5 10 15 Ala Leu Ala Ala Pro Val Asn Thr Thr Thr Glu Asp
Glu Thr Ala Gln 20 25 30 Ile Pro Ala Glu Ala Val Ile Gly Tyr Ser
Asp Leu Glu Gly Asp Phe 35 40 45 Asp Val Ala Val Leu Pro Phe Ser
Asn Ser Thr Asn Asn Gly Leu Leu 50 55 60 Phe Ile Asn Thr Thr Ile
Ala Ser Ile Ala Ala Lys Glu Glu Gly Val 65 70 75 80 Ser Leu Glu Lys
Arg Glu Ala Glu Ala Glu Phe His His His His His 85 90 95 His Ala
Gly Ile His Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu 100 105 110
Ala Val Ser Leu Gly Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Lys 115
120 125 Ser Val Ser Thr Ser Ser Tyr Ser Tyr Met His Trp Tyr Gln Gln
Lys 130 135 140 Pro Gly Gln Pro Pro Lys Leu Leu Ile Lys Tyr Ala Ser
Tyr Leu Glu 145 150 155 160 Ser Gly Val Pro Ala Arg Phe Ser Gly Ser
Gly Ser Gly Thr Asp Phe 165 170 175 Thr Leu Asn Ile His Pro Val Glu
Glu Glu Asp Ala Ala Thr Tyr Tyr 180 185 190 Cys Gln His Ser Arg Glu
Phe Pro Trp Thr Phe Gly Gly Gly Thr Lys 195 200 205 Leu Glu Ile Lys
Arg Ala Asp Ala Ala Pro Gly Gly Gly Gly Ser Gly 210 215 220 Gly Gly
Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser 225 230 235
240 Gly Gly Gly Leu Val Lys Pro Gly Gly Ser Arg Lys Leu Ser Cys Ala
245 250 255 Ala Ser Gly Phe Thr Phe Ser Asn Tyr Gly Met His Trp Val
Arg Gln 260 265 270 Ala Pro Glu Lys Gly Leu Glu Trp Val Ala Tyr Ile
Ser Ser Gly Ser 275 280 285 Ser Thr Ile Tyr Tyr Ala Asp Thr Val Lys
Gly Arg Phe Thr Ile Ser 290 295 300 Arg Asp Asn Ala Lys Asn Thr Leu
Phe Leu Gln Met Thr Ser Leu Arg 305 310 315 320 Ser Glu Asp Thr Ala
Met Arg Gly Leu Leu Leu Asp Tyr Trp Gly Gln 325 330 335 Gly Thr Thr
Leu Thr Val Ser Ser Leu Glu Gln Lys Leu Ile Ser Glu 340 345 350 Glu
Asp Leu Asn Ser Ala Val Asp Ala Ser Thr Lys Gly Pro Ser Val 355 360
365 Phe Pro Leu Ala Pro Leu Glu Ser Ser Gly Ser Met Thr Glu Arg Arg
370 375 380 Val Pro Phe Ser Leu Leu Arg Gly Pro Ser Trp Asp Pro Phe
Arg Asp 385 390 395 400 Trp Tyr Pro His Ser Arg Leu Phe Asp Gln Ala
Phe Gly Leu Pro Arg 405 410 415 Leu Pro Glu Glu Trp Ser Gln Trp Leu
Gly Gly Ser Ser Trp Pro Gly 420 425 430 Tyr Val Arg Pro Leu Pro Pro
Ala Ala Ile Glu Ser Pro Ala Val Ala 435 440 445 Ala Pro Ala Tyr Ser
Arg Ala Leu Ser Arg Gln Leu Ser Ser Gly Val 450 455 460 Ser Glu Ile
Arg His Thr Ala Asp Arg Trp Arg Val Ser Leu Asp Val 465 470 475 480
Asn His Phe Ala Pro Asp Glu Leu Thr Val Lys Thr Lys Asp Gly Val 485
490 495 Val Glu Ile Thr Gly Lys His Glu Glu Arg Gln Asp Glu His Gly
Tyr 500 505 510 Ile Ser Arg Cys Phe Thr Arg Lys Tyr Thr Leu Pro Pro
Gly Val Asp 515 520 525 Pro Thr Gln Val Ser Ser Ser Leu Ser Pro Glu
Gly Thr Leu Thr Val 530 535 540 Glu Ala Pro Met Pro Lys Leu Ala Thr
Gln Ser Asn Glu Ile Thr Ile 545 550 555 560 Pro Val Thr Phe Glu Ser
Arg Ala Gln Leu Gly Gly Pro Glu Ala Ala 565 570 575 Lys Ser Asp Glu
Thr Ala Ala Lys 580 283069DNAArtificial Sequence3E10 Fv-Hsp70
nucleotide sequence encoding a chimeric protein comprising a single
chain Fv antibody fragment derived from Mus musculus 3E10 hybridoma
and Hsp70 from Homo sapiens joined by human CH1 linker and swivel
sequence 28atg aga ttt cct tca ctt ttt act gct gtt tta ttc gca gca
tcc tcc 48Met Arg Phe Pro Ser Leu Phe Thr Ala Val Leu Phe Ala Ala
Ser Ser 1 5 10 15 gca tta gct gct cca gtc aac act aca aca gaa gat
gaa acg gca caa 96Ala Leu Ala Ala Pro Val Asn Thr Thr Thr Glu Asp
Glu Thr Ala Gln 20 25 30 att ccg gct gaa gct gtc atc ggt tac tca
gat tta gaa ggg gat ttc 144Ile Pro Ala Glu Ala Val Ile Gly Tyr Ser
Asp Leu Glu Gly Asp Phe 35 40 45 gat gtt gct gtt ttg cca ttt tcc
aac agc aca aat aac ggg tta ttg 192Asp Val Ala Val Leu Pro Phe Ser
Asn Ser Thr Asn Asn Gly Leu Leu 50 55 60 ttt ata aat act act att
gcc agc att gct gct aaa gaa gaa ggg gta 240Phe Ile Asn Thr Thr Ile
Ala Ser Ile Ala Ala Lys Glu Glu Gly Val 65 70 75 80 tct ctc gag aaa
aga gag gct gaa gct gaa ttc cat cac cat cac cat 288Ser Leu Glu Lys
Arg Glu Ala Glu Ala Glu Phe His His His His His 85 90 95 cac gca
ggg att cac gac att gtc ctg aca cag tct cct gct tcc tta 336His Ala
Gly Ile His Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu 100 105 110
gct gta tct ctg ggg cag agg gcc acc atc tcc tgc agg gcc agc aaa
384Ala Val Ser Leu Gly Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Lys
115 120 125 agt gtc agt aca tct agc tat agt tac atg cac tgg tac caa
cag aaa 432Ser Val Ser Thr Ser Ser Tyr Ser Tyr Met His Trp Tyr Gln
Gln Lys 130 135 140 cca gga cag cca ccc aaa ctc ctc atc aag tat gca
tcc tac cta gaa 480Pro Gly Gln Pro Pro Lys Leu Leu Ile Lys Tyr Ala
Ser Tyr Leu Glu 145 150 155 160 tct ggg gtt cct gcc agg ttc agt ggc
agt ggg tct ggg aca gac ttc 528Ser Gly Val Pro Ala Arg Phe Ser Gly
Ser Gly Ser Gly Thr Asp Phe 165 170 175 acc ctc aac atc cat cct gtg
gag gag gag gat gct gca aca tat tac 576Thr Leu Asn Ile His Pro Val
Glu Glu Glu Asp Ala Ala Thr Tyr Tyr 180 185 190 tgt cag cac agt agg
gag ttt ccg tgg acg ttc ggt gga ggc acc aag 624Cys Gln His Ser Arg
Glu Phe Pro Trp Thr Phe Gly Gly Gly Thr Lys 195 200 205 ctg gaa atc
aaa cgg gct gat gct gca ccc ggg ggt ggc ggt tct ggc 672Leu Glu Ile
Lys Arg Ala Asp Ala Ala Pro Gly Gly Gly Gly Ser Gly 210 215 220 ggt
ggc ggt tct gga ggc ggt ggc tct gag gtg cag ctg gtg gag tct 720Gly
Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser 225 230
235 240 ggg gga ggc tta gtg aag cct gga ggg tcc cgg aaa ctc tcc tgt
gca 768Gly Gly Gly Leu Val Lys Pro Gly Gly Ser Arg Lys Leu Ser Cys
Ala 245 250 255 gcc tct gga ttc act ttc agt aac tat gga atg cac tgg
gtc cgt cag 816Ala Ser Gly Phe Thr Phe Ser Asn Tyr Gly Met His Trp
Val Arg Gln 260 265 270 gct cca gag aag ggg ctg gag tgg gtt gca tac
att agt agt ggc agt 864Ala Pro Glu Lys Gly Leu Glu Trp Val Ala Tyr
Ile Ser Ser Gly Ser 275 280 285 agt acc atc tac tat gca gac aca gtg
aag ggc cga ttc acc atc tcc 912Ser Thr Ile Tyr Tyr Ala Asp Thr Val
Lys Gly Arg Phe Thr Ile Ser 290 295 300 aga gac aat gcc aag aac acc
ctg ttc ctg caa atg acc agt cta agg 960Arg Asp Asn Ala Lys Asn Thr
Leu Phe Leu Gln Met Thr Ser Leu Arg 305 310 315 320 tct gag gac aca
gcc atg cgg ggg tta cta ctt gac tac tgg ggc caa 1008Ser Glu Asp Thr
Ala Met Arg Gly Leu Leu Leu Asp Tyr Trp Gly Gln 325 330 335 ggc acc
act ctc aca gtc tcc tca cta gaa caa aaa ctc atc tca gaa 1056Gly Thr
Thr Leu Thr Val Ser Ser Leu Glu Gln Lys Leu Ile Ser Glu 340 345 350
gag gat ctg aat agc gcc gtc gac gct tcc acc aag ggc cca tcc gtc
1104Glu Asp Leu Asn Ser Ala Val Asp Ala Ser Thr Lys Gly Pro Ser Val
355 360 365 ttc ccc ctg gcg ccc ctg gag tct tcc gga tcc atg gcc aaa
gcc gcg 1152Phe Pro Leu Ala Pro Leu Glu Ser Ser Gly Ser Met Ala Lys
Ala Ala 370 375 380 gcg atc ggc atc gac ctg ggc acc acc tac tcc tgc
gtg ggg gtg ttc 1200Ala Ile Gly Ile Asp Leu Gly Thr Thr Tyr Ser Cys
Val Gly Val Phe 385 390 395 400 caa cac ggc aag gtg gag atc atc gcc
aac gac cag ggc aac cgc acc 1248Gln His Gly Lys Val Glu Ile Ile Ala
Asn Asp Gln Gly Asn Arg Thr 405 410 415 acc ccc agc tac gtg gcc ttc
acg gac acc gag cgg ctc atc ggg gat 1296Thr Pro Ser Tyr Val Ala Phe
Thr Asp Thr Glu Arg Leu Ile Gly Asp 420 425 430 gcg gcc aag aac cag
gtg gcg ctg aac ccg cag aac acc gtg ttt gac 1344Ala Ala Lys Asn Gln
Val Ala Leu Asn Pro Gln Asn Thr Val Phe Asp 435 440 445 gcg aag cgg
ctg atc ggc cgc aag ttc ggc gac ccg gtg gtg cag tcg 1392Ala Lys Arg
Leu Ile Gly Arg Lys Phe Gly Asp Pro Val Val Gln Ser 450 455 460 gac
atg aag cac tgg cct ttc cag gtg atc aac gac gga gac aag ccc 1440Asp
Met Lys His Trp Pro Phe Gln Val Ile Asn Asp Gly Asp Lys Pro 465 470
475 480 aag gtg cag gtg agc tac aag ggg gag acc aag gca ttc tac ccc
gag 1488Lys Val Gln Val Ser Tyr Lys Gly Glu Thr Lys Ala Phe Tyr Pro
Glu 485 490 495 gag atc tcg tcc atg gtg ctg acc aag atg aag gag atc
gcc gag gcg 1536Glu Ile Ser Ser Met Val Leu Thr Lys Met Lys Glu Ile
Ala Glu Ala 500 505 510 tac ctg ggc tac ccg gtg acc aac gcg gtg atc
acc gtg ccg gcc tac 1584Tyr Leu Gly Tyr Pro Val Thr Asn Ala Val Ile
Thr Val Pro Ala Tyr 515 520 525 ttc aac gac tcg cag cgc cag gcc acc
aag gat gcg ggt gtg atc gcg 1632Phe Asn Asp Ser Gln Arg Gln Ala Thr
Lys Asp Ala Gly Val Ile Ala 530 535 540 ggg ctc aac gtg ctg cgg atc
atc aac gag ccc acg gcc gcc gcc atc 1680Gly Leu Asn Val Leu Arg Ile
Ile Asn Glu Pro Thr Ala Ala Ala Ile 545 550 555 560 gcc tac ggc ctg
gac aga acg ggc aag ggg gag cgc aac gtg ctc atc 1728Ala Tyr Gly Leu
Asp Arg Thr Gly Lys Gly Glu Arg Asn Val Leu Ile 565 570 575 ttt gac
ctg ggc ggg ggc acc ttc gac gtg tcc atc ctg acg atc gac 1776Phe Asp
Leu Gly Gly Gly Thr Phe Asp Val Ser Ile Leu Thr Ile Asp 580 585 590
gac ggc atc ttc gag gtg aag gcc acg gcc ggg gac acc cac ctg ggt
1824Asp Gly Ile Phe Glu Val Lys Ala Thr Ala Gly Asp Thr His Leu Gly
595 600 605 ggg gag gac ttt gac aac agg ctg gtg aac cac ttc gtg gag
gag ttc 1872Gly Glu Asp Phe Asp Asn Arg Leu Val Asn His Phe Val Glu
Glu Phe 610 615 620 aag aga aaa cac aag aag gac atc agc cag aac aag
cga gcc gtg agg 1920Lys Arg Lys His Lys Lys Asp Ile Ser Gln Asn Lys
Arg Ala Val Arg 625 630 635 640 cgg ctg cgc acc gcc tgc gag agg gcc
aag agg acc ctg tcg tcc agc 1968Arg Leu Arg Thr Ala Cys Glu Arg Ala
Lys Arg Thr Leu Ser Ser Ser 645 650 655 acc cag gcc agc ctg gag atc
gac tcc ctg ttt gag ggc atc gac ttc 2016Thr Gln Ala Ser Leu Glu Ile
Asp Ser Leu Phe Glu Gly Ile Asp Phe 660 665 670 tac acg tcc atc acc
agg gcg agg ttc gag gag ctg tgc tcc gac ctg 2064Tyr Thr Ser Ile Thr
Arg Ala Arg Phe Glu Glu Leu Cys Ser Asp Leu 675 680 685 ttc cga agc
acc ctg gag ccc gtg gag aag gct ctg cgc gac gcc aag 2112Phe Arg Ser
Thr Leu Glu Pro Val Glu Lys Ala Leu Arg Asp Ala Lys 690 695 700 ctg
gac aag gcc cag att cac gac ctg gtc ctg gtc ggg ggc tcc acc 2160Leu
Asp Lys Ala Gln Ile His Asp Leu Val Leu Val Gly Gly Ser Thr 705 710
715 720 cgc atc ccc aag gtg cag aag ctg ctg cag gac ttc ttc aac ggg
cgc 2208Arg Ile Pro Lys Val Gln Lys Leu Leu Gln Asp Phe Phe Asn Gly
Arg 725 730 735 gac ctg aac aag agc atc aac ccc gac gag gct gtg gcc
tac ggg gcg 2256Asp Leu Asn Lys Ser Ile Asn Pro Asp Glu Ala Val Ala
Tyr Gly Ala 740 745 750 gcg gtg cag gcg gcc atc ctg atg ggg gac aag
tcc gag aac gtg cag 2304Ala Val Gln Ala Ala Ile Leu Met Gly Asp Lys
Ser Glu Asn Val Gln 755 760 765 gac ctg ctg ctg ctg gac gtg gct ccc
ctg tcg ctg ggg ctg gag acg 2352Asp Leu Leu Leu Leu Asp Val Ala Pro
Leu Ser Leu Gly Leu Glu Thr 770 775 780 gcc gga ggc gtg atg act gcc
ctg atc aag cgc aac tcc acc atc ccc 2400Ala Gly Gly Val Met Thr Ala
Leu Ile Lys Arg Asn Ser Thr Ile Pro 785 790 795 800 acc aag cag acg
cag atc ttc acc acc tac tcc gac aac caa ccc ggg 2448Thr Lys Gln Thr
Gln Ile Phe Thr Thr Tyr Ser Asp Asn Gln Pro Gly 805 810 815 gtg ctg
atc cag gtg tac gag ggc gag agg gcc atg acg aaa gac aac 2496Val Leu
Ile Gln Val Tyr Glu Gly Glu Arg Ala Met Thr Lys Asp Asn 820 825
830
aat ctg ttg ggg cgc ttc gag ctg agc ggc atc cct ccg gcc ccc agg
2544Asn Leu Leu Gly Arg Phe Glu Leu Ser Gly Ile Pro Pro Ala Pro Arg
835 840 845 ggc gtg ccc cag atc gag gtg acc ttc gac atc gat gcc aac
ggc atc 2592Gly Val Pro Gln Ile Glu Val Thr Phe Asp Ile Asp Ala Asn
Gly Ile 850 855 860 ctg aac gtc acg gcc acg gac aag agc acc ggc aag
gcc aac aag atc 2640Leu Asn Val Thr Ala Thr Asp Lys Ser Thr Gly Lys
Ala Asn Lys Ile 865 870 875 880 acc atc acc aac gac aag ggc cgc ctg
agc aag gag gag atc gag cgc 2688Thr Ile Thr Asn Asp Lys Gly Arg Leu
Ser Lys Glu Glu Ile Glu Arg 885 890 895 atg gtg cag gag gcg gag aag
tac aaa gcg gag gac gag gtg cag cgc 2736Met Val Gln Glu Ala Glu Lys
Tyr Lys Ala Glu Asp Glu Val Gln Arg 900 905 910 gag agg gtg tca gcc
aag aac gcc ctg gag tcc tac gcc ttc aac atg 2784Glu Arg Val Ser Ala
Lys Asn Ala Leu Glu Ser Tyr Ala Phe Asn Met 915 920 925 aag agc gcc
gtg gag gat gag ggg ctc aag ggc aag atc agc gag gcg 2832Lys Ser Ala
Val Glu Asp Glu Gly Leu Lys Gly Lys Ile Ser Glu Ala 930 935 940 gac
aag aag aag gtt ctg gac aag tgt caa gag gtc atc tcg tgg ctg 2880Asp
Lys Lys Lys Val Leu Asp Lys Cys Gln Glu Val Ile Ser Trp Leu 945 950
955 960 gac gcc aac acc ttg gcc gag aag gac gag ttt gag cac aag agg
aag 2928Asp Ala Asn Thr Leu Ala Glu Lys Asp Glu Phe Glu His Lys Arg
Lys 965 970 975 gag ctg gag cag gtg tgt aac ccc atc atc agc gga ctg
tac cag ggt 2976Glu Leu Glu Gln Val Cys Asn Pro Ile Ile Ser Gly Leu
Tyr Gln Gly 980 985 990 gcc ggt ggt ccc ggg cct ggc ggc ttc ggg gct
cag ggt ccc aag gga 3024Ala Gly Gly Pro Gly Pro Gly Gly Phe Gly Ala
Gln Gly Pro Lys Gly 995 1000 1005 ggg tct ggg tca ggc cct acc att
gag gag gtg gat tag tctaga 3069Gly Ser Gly Ser Gly Pro Thr Ile Glu
Glu Val Asp 1010 1015 1020 291020PRTArtificial SequenceSynthetic
Construct 29Met Arg Phe Pro Ser Leu Phe Thr Ala Val Leu Phe Ala Ala
Ser Ser 1 5 10 15 Ala Leu Ala Ala Pro Val Asn Thr Thr Thr Glu Asp
Glu Thr Ala Gln 20 25 30 Ile Pro Ala Glu Ala Val Ile Gly Tyr Ser
Asp Leu Glu Gly Asp Phe 35 40 45 Asp Val Ala Val Leu Pro Phe Ser
Asn Ser Thr Asn Asn Gly Leu Leu 50 55 60 Phe Ile Asn Thr Thr Ile
Ala Ser Ile Ala Ala Lys Glu Glu Gly Val 65 70 75 80 Ser Leu Glu Lys
Arg Glu Ala Glu Ala Glu Phe His His His His His 85 90 95 His Ala
Gly Ile His Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu 100 105 110
Ala Val Ser Leu Gly Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Lys 115
120 125 Ser Val Ser Thr Ser Ser Tyr Ser Tyr Met His Trp Tyr Gln Gln
Lys 130 135 140 Pro Gly Gln Pro Pro Lys Leu Leu Ile Lys Tyr Ala Ser
Tyr Leu Glu 145 150 155 160 Ser Gly Val Pro Ala Arg Phe Ser Gly Ser
Gly Ser Gly Thr Asp Phe 165 170 175 Thr Leu Asn Ile His Pro Val Glu
Glu Glu Asp Ala Ala Thr Tyr Tyr 180 185 190 Cys Gln His Ser Arg Glu
Phe Pro Trp Thr Phe Gly Gly Gly Thr Lys 195 200 205 Leu Glu Ile Lys
Arg Ala Asp Ala Ala Pro Gly Gly Gly Gly Ser Gly 210 215 220 Gly Gly
Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser 225 230 235
240 Gly Gly Gly Leu Val Lys Pro Gly Gly Ser Arg Lys Leu Ser Cys Ala
245 250 255 Ala Ser Gly Phe Thr Phe Ser Asn Tyr Gly Met His Trp Val
Arg Gln 260 265 270 Ala Pro Glu Lys Gly Leu Glu Trp Val Ala Tyr Ile
Ser Ser Gly Ser 275 280 285 Ser Thr Ile Tyr Tyr Ala Asp Thr Val Lys
Gly Arg Phe Thr Ile Ser 290 295 300 Arg Asp Asn Ala Lys Asn Thr Leu
Phe Leu Gln Met Thr Ser Leu Arg 305 310 315 320 Ser Glu Asp Thr Ala
Met Arg Gly Leu Leu Leu Asp Tyr Trp Gly Gln 325 330 335 Gly Thr Thr
Leu Thr Val Ser Ser Leu Glu Gln Lys Leu Ile Ser Glu 340 345 350 Glu
Asp Leu Asn Ser Ala Val Asp Ala Ser Thr Lys Gly Pro Ser Val 355 360
365 Phe Pro Leu Ala Pro Leu Glu Ser Ser Gly Ser Met Ala Lys Ala Ala
370 375 380 Ala Ile Gly Ile Asp Leu Gly Thr Thr Tyr Ser Cys Val Gly
Val Phe 385 390 395 400 Gln His Gly Lys Val Glu Ile Ile Ala Asn Asp
Gln Gly Asn Arg Thr 405 410 415 Thr Pro Ser Tyr Val Ala Phe Thr Asp
Thr Glu Arg Leu Ile Gly Asp 420 425 430 Ala Ala Lys Asn Gln Val Ala
Leu Asn Pro Gln Asn Thr Val Phe Asp 435 440 445 Ala Lys Arg Leu Ile
Gly Arg Lys Phe Gly Asp Pro Val Val Gln Ser 450 455 460 Asp Met Lys
His Trp Pro Phe Gln Val Ile Asn Asp Gly Asp Lys Pro 465 470 475 480
Lys Val Gln Val Ser Tyr Lys Gly Glu Thr Lys Ala Phe Tyr Pro Glu 485
490 495 Glu Ile Ser Ser Met Val Leu Thr Lys Met Lys Glu Ile Ala Glu
Ala 500 505 510 Tyr Leu Gly Tyr Pro Val Thr Asn Ala Val Ile Thr Val
Pro Ala Tyr 515 520 525 Phe Asn Asp Ser Gln Arg Gln Ala Thr Lys Asp
Ala Gly Val Ile Ala 530 535 540 Gly Leu Asn Val Leu Arg Ile Ile Asn
Glu Pro Thr Ala Ala Ala Ile 545 550 555 560 Ala Tyr Gly Leu Asp Arg
Thr Gly Lys Gly Glu Arg Asn Val Leu Ile 565 570 575 Phe Asp Leu Gly
Gly Gly Thr Phe Asp Val Ser Ile Leu Thr Ile Asp 580 585 590 Asp Gly
Ile Phe Glu Val Lys Ala Thr Ala Gly Asp Thr His Leu Gly 595 600 605
Gly Glu Asp Phe Asp Asn Arg Leu Val Asn His Phe Val Glu Glu Phe 610
615 620 Lys Arg Lys His Lys Lys Asp Ile Ser Gln Asn Lys Arg Ala Val
Arg 625 630 635 640 Arg Leu Arg Thr Ala Cys Glu Arg Ala Lys Arg Thr
Leu Ser Ser Ser 645 650 655 Thr Gln Ala Ser Leu Glu Ile Asp Ser Leu
Phe Glu Gly Ile Asp Phe 660 665 670 Tyr Thr Ser Ile Thr Arg Ala Arg
Phe Glu Glu Leu Cys Ser Asp Leu 675 680 685 Phe Arg Ser Thr Leu Glu
Pro Val Glu Lys Ala Leu Arg Asp Ala Lys 690 695 700 Leu Asp Lys Ala
Gln Ile His Asp Leu Val Leu Val Gly Gly Ser Thr 705 710 715 720 Arg
Ile Pro Lys Val Gln Lys Leu Leu Gln Asp Phe Phe Asn Gly Arg 725 730
735 Asp Leu Asn Lys Ser Ile Asn Pro Asp Glu Ala Val Ala Tyr Gly Ala
740 745 750 Ala Val Gln Ala Ala Ile Leu Met Gly Asp Lys Ser Glu Asn
Val Gln 755 760 765 Asp Leu Leu Leu Leu Asp Val Ala Pro Leu Ser Leu
Gly Leu Glu Thr 770 775 780 Ala Gly Gly Val Met Thr Ala Leu Ile Lys
Arg Asn Ser Thr Ile Pro 785 790 795 800 Thr Lys Gln Thr Gln Ile Phe
Thr Thr Tyr Ser Asp Asn Gln Pro Gly 805 810 815 Val Leu Ile Gln Val
Tyr Glu Gly Glu Arg Ala Met Thr Lys Asp Asn 820 825 830 Asn Leu Leu
Gly Arg Phe Glu Leu Ser Gly Ile Pro Pro Ala Pro Arg 835 840 845 Gly
Val Pro Gln Ile Glu Val Thr Phe Asp Ile Asp Ala Asn Gly Ile 850 855
860 Leu Asn Val Thr Ala Thr Asp Lys Ser Thr Gly Lys Ala Asn Lys Ile
865 870 875 880 Thr Ile Thr Asn Asp Lys Gly Arg Leu Ser Lys Glu Glu
Ile Glu Arg 885 890 895 Met Val Gln Glu Ala Glu Lys Tyr Lys Ala Glu
Asp Glu Val Gln Arg 900 905 910 Glu Arg Val Ser Ala Lys Asn Ala Leu
Glu Ser Tyr Ala Phe Asn Met 915 920 925 Lys Ser Ala Val Glu Asp Glu
Gly Leu Lys Gly Lys Ile Ser Glu Ala 930 935 940 Asp Lys Lys Lys Val
Leu Asp Lys Cys Gln Glu Val Ile Ser Trp Leu 945 950 955 960 Asp Ala
Asn Thr Leu Ala Glu Lys Asp Glu Phe Glu His Lys Arg Lys 965 970 975
Glu Leu Glu Gln Val Cys Asn Pro Ile Ile Ser Gly Leu Tyr Gln Gly 980
985 990 Ala Gly Gly Pro Gly Pro Gly Gly Phe Gly Ala Gln Gly Pro Lys
Gly 995 1000 1005 Gly Ser Gly Ser Gly Pro Thr Ile Glu Glu Val Asp
1010 1015 1020 301020PRTArtificial Sequence3E10 Fv-Hsp70 amino acid
sequence for a chimeric protein comprising a single chain Fv
antibody fragment derived from Mus musculus 3E10 hybridoma and
Hsp70 from Homo sapiens joined by human CH1 linker and swivel
sequence 30Met Arg Phe Pro Ser Leu Phe Thr Ala Val Leu Phe Ala Ala
Ser Ser 1 5 10 15 Ala Leu Ala Ala Pro Val Asn Thr Thr Thr Glu Asp
Glu Thr Ala Gln 20 25 30 Ile Pro Ala Glu Ala Val Ile Gly Tyr Ser
Asp Leu Glu Gly Asp Phe 35 40 45 Asp Val Ala Val Leu Pro Phe Ser
Asn Ser Thr Asn Asn Gly Leu Leu 50 55 60 Phe Ile Asn Thr Thr Ile
Ala Ser Ile Ala Ala Lys Glu Glu Gly Val 65 70 75 80 Ser Leu Glu Lys
Arg Glu Ala Glu Ala Glu Phe His His His His His 85 90 95 His Ala
Gly Ile His Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu 100 105 110
Ala Val Ser Leu Gly Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Lys 115
120 125 Ser Val Ser Thr Ser Ser Tyr Ser Tyr Met His Trp Tyr Gln Gln
Lys 130 135 140 Pro Gly Gln Pro Pro Lys Leu Leu Ile Lys Tyr Ala Ser
Tyr Leu Glu 145 150 155 160 Ser Gly Val Pro Ala Arg Phe Ser Gly Ser
Gly Ser Gly Thr Asp Phe 165 170 175 Thr Leu Asn Ile His Pro Val Glu
Glu Glu Asp Ala Ala Thr Tyr Tyr 180 185 190 Cys Gln His Ser Arg Glu
Phe Pro Trp Thr Phe Gly Gly Gly Thr Lys 195 200 205 Leu Glu Ile Lys
Arg Ala Asp Ala Ala Pro Gly Gly Gly Gly Ser Gly 210 215 220 Gly Gly
Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser 225 230 235
240 Gly Gly Gly Leu Val Lys Pro Gly Gly Ser Arg Lys Leu Ser Cys Ala
245 250 255 Ala Ser Gly Phe Thr Phe Ser Asn Tyr Gly Met His Trp Val
Arg Gln 260 265 270 Ala Pro Glu Lys Gly Leu Glu Trp Val Ala Tyr Ile
Ser Ser Gly Ser 275 280 285 Ser Thr Ile Tyr Tyr Ala Asp Thr Val Lys
Gly Arg Phe Thr Ile Ser 290 295 300 Arg Asp Asn Ala Lys Asn Thr Leu
Phe Leu Gln Met Thr Ser Leu Arg 305 310 315 320 Ser Glu Asp Thr Ala
Met Arg Gly Leu Leu Leu Asp Tyr Trp Gly Gln 325 330 335 Gly Thr Thr
Leu Thr Val Ser Ser Leu Glu Gln Lys Leu Ile Ser Glu 340 345 350 Glu
Asp Leu Asn Ser Ala Val Asp Ala Ser Thr Lys Gly Pro Ser Val 355 360
365 Phe Pro Leu Ala Pro Leu Glu Ser Ser Gly Ser Met Ala Lys Ala Ala
370 375 380 Ala Ile Gly Ile Asp Leu Gly Thr Thr Tyr Ser Cys Val Gly
Val Phe 385 390 395 400 Gln His Gly Lys Val Glu Ile Ile Ala Asn Asp
Gln Gly Asn Arg Thr 405 410 415 Thr Pro Ser Tyr Val Ala Phe Thr Asp
Thr Glu Arg Leu Ile Gly Asp 420 425 430 Ala Ala Lys Asn Gln Val Ala
Leu Asn Pro Gln Asn Thr Val Phe Asp 435 440 445 Ala Lys Arg Leu Ile
Gly Arg Lys Phe Gly Asp Pro Val Val Gln Ser 450 455 460 Asp Met Lys
His Trp Pro Phe Gln Val Ile Asn Asp Gly Asp Lys Pro 465 470 475 480
Lys Val Gln Val Ser Tyr Lys Gly Glu Thr Lys Ala Phe Tyr Pro Glu 485
490 495 Glu Ile Ser Ser Met Val Leu Thr Lys Met Lys Glu Ile Ala Glu
Ala 500 505 510 Tyr Leu Gly Tyr Pro Val Thr Asn Ala Val Ile Thr Val
Pro Ala Tyr 515 520 525 Phe Asn Asp Ser Gln Arg Gln Ala Thr Lys Asp
Ala Gly Val Ile Ala 530 535 540 Gly Leu Asn Val Leu Arg Ile Ile Asn
Glu Pro Thr Ala Ala Ala Ile 545 550 555 560 Ala Tyr Gly Leu Asp Arg
Thr Gly Lys Gly Glu Arg Asn Val Leu Ile 565 570 575 Phe Asp Leu Gly
Gly Gly Thr Phe Asp Val Ser Ile Leu Thr Ile Asp 580 585 590 Asp Gly
Ile Phe Glu Val Lys Ala Thr Ala Gly Asp Thr His Leu Gly 595 600 605
Gly Glu Asp Phe Asp Asn Arg Leu Val Asn His Phe Val Glu Glu Phe 610
615 620 Lys Arg Lys His Lys Lys Asp Ile Ser Gln Asn Lys Arg Ala Val
Arg 625 630 635 640 Arg Leu Arg Thr Ala Cys Glu Arg Ala Lys Arg Thr
Leu Ser Ser Ser 645 650 655 Thr Gln Ala Ser Leu Glu Ile Asp Ser Leu
Phe Glu Gly Ile Asp Phe 660 665 670 Tyr Thr Ser Ile Thr Arg Ala Arg
Phe Glu Glu Leu Cys Ser Asp Leu 675 680 685 Phe Arg Ser Thr Leu Glu
Pro Val Glu Lys Ala Leu Arg Asp Ala Lys 690 695 700 Leu Asp Lys Ala
Gln Ile His Asp Leu Val Leu Val Gly Gly Ser Thr 705 710 715 720 Arg
Ile Pro Lys Val Gln Lys Leu Leu Gln Asp Phe Phe Asn Gly Arg 725 730
735 Asp Leu Asn Lys Ser Ile Asn Pro Asp Glu Ala Val Ala Tyr Gly Ala
740 745 750 Ala Val Gln Ala Ala Ile Leu Met Gly Asp Lys Ser Glu Asn
Val Gln 755 760 765 Asp Leu Leu Leu Leu Asp Val Ala Pro Leu Ser Leu
Gly Leu Glu Thr 770 775 780 Ala Gly Gly Val Met Thr Ala Leu Ile Lys
Arg Asn Ser Thr Ile Pro 785 790 795 800 Thr Lys Gln Thr Gln Ile Phe
Thr Thr Tyr Ser Asp Asn Gln Pro Gly 805 810 815 Val Leu Ile Gln Val
Tyr Glu Gly Glu Arg Ala Met Thr Lys Asp Asn 820 825 830 Asn Leu Leu
Gly Arg Phe Glu Leu Ser Gly Ile Pro Pro Ala Pro Arg 835 840 845 Gly
Val Pro Gln Ile Glu Val Thr Phe Asp Ile Asp Ala Asn Gly Ile 850 855
860 Leu Asn Val Thr Ala Thr Asp Lys Ser Thr Gly Lys Ala Asn Lys Ile
865 870 875 880 Thr Ile Thr Asn Asp Lys Gly Arg Leu Ser Lys Glu Glu
Ile Glu Arg 885 890 895 Met Val Gln Glu Ala Glu Lys Tyr Lys Ala Glu
Asp Glu Val Gln Arg 900 905 910 Glu
Arg Val Ser Ala Lys Asn Ala Leu Glu Ser Tyr Ala Phe Asn Met 915 920
925 Lys Ser Ala Val Glu Asp Glu Gly Leu Lys Gly Lys Ile Ser Glu Ala
930 935 940 Asp Lys Lys Lys Val Leu Asp Lys Cys Gln Glu Val Ile Ser
Trp Leu 945 950 955 960 Asp Ala Asn Thr Leu Ala Glu Lys Asp Glu Phe
Glu His Lys Arg Lys 965 970 975 Glu Leu Glu Gln Val Cys Asn Pro Ile
Ile Ser Gly Leu Tyr Gln Gly 980 985 990 Ala Gly Gly Pro Gly Pro Gly
Gly Phe Gly Ala Gln Gly Pro Lys Gly 995 1000 1005 Gly Ser Gly Ser
Gly Pro Thr Ile Glu Glu Val Asp 1010 1015 1020 313108DNAArtificial
Sequence3E10 Fv-GRP78 nucleotide sequence encoding a chimeric
protein comprising a single chain Fv antibody fragment derived from
Mus musculus 3E10 hybridoma and GRP78 from Homo sapiens joined by
human CH1 linker and swivel sequence 31atg aga ttt cct tca ctt ttt
act gct gtt tta ttc gca gca tcc tcc 48Met Arg Phe Pro Ser Leu Phe
Thr Ala Val Leu Phe Ala Ala Ser Ser 1 5 10 15 gca tta gct gct cca
gtc aac act aca aca gaa gat gaa acg gca caa 96Ala Leu Ala Ala Pro
Val Asn Thr Thr Thr Glu Asp Glu Thr Ala Gln 20 25 30 att ccg gct
gaa gct gtc atc ggt tac tca gat tta gaa ggg gat ttc 144Ile Pro Ala
Glu Ala Val Ile Gly Tyr Ser Asp Leu Glu Gly Asp Phe 35 40 45 gat
gtt gct gtt ttg cca ttt tcc aac agc aca aat aac ggg tta ttg 192Asp
Val Ala Val Leu Pro Phe Ser Asn Ser Thr Asn Asn Gly Leu Leu 50 55
60 ttt ata aat act act att gcc agc att gct gct aaa gaa gaa ggg gta
240Phe Ile Asn Thr Thr Ile Ala Ser Ile Ala Ala Lys Glu Glu Gly Val
65 70 75 80 tct ctc gag aaa aga gag gct gaa gct gaa ttc cat cac cat
cac cat 288Ser Leu Glu Lys Arg Glu Ala Glu Ala Glu Phe His His His
His His 85 90 95 cac gca ggg att cac gac att gtc ctg aca cag tct
cct gct tcc tta 336His Ala Gly Ile His Asp Ile Val Leu Thr Gln Ser
Pro Ala Ser Leu 100 105 110 gct gta tct ctg ggg cag agg gcc acc atc
tcc tgc agg gcc agc aaa 384Ala Val Ser Leu Gly Gln Arg Ala Thr Ile
Ser Cys Arg Ala Ser Lys 115 120 125 agt gtc agt aca tct agc tat agt
tac atg cac tgg tac caa cag aaa 432Ser Val Ser Thr Ser Ser Tyr Ser
Tyr Met His Trp Tyr Gln Gln Lys 130 135 140 cca gga cag cca ccc aaa
ctc ctc atc aag tat gca tcc tac cta gaa 480Pro Gly Gln Pro Pro Lys
Leu Leu Ile Lys Tyr Ala Ser Tyr Leu Glu 145 150 155 160 tct ggg gtt
cct gcc agg ttc agt ggc agt ggg tct ggg aca gac ttc 528Ser Gly Val
Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe 165 170 175 acc
ctc aac atc cat cct gtg gag gag gag gat gct gca aca tat tac 576Thr
Leu Asn Ile His Pro Val Glu Glu Glu Asp Ala Ala Thr Tyr Tyr 180 185
190 tgt cag cac agt agg gag ttt ccg tgg acg ttc ggt gga ggc acc aag
624Cys Gln His Ser Arg Glu Phe Pro Trp Thr Phe Gly Gly Gly Thr Lys
195 200 205 ctg gaa atc aaa cgg gct gat gct gca ccc ggg ggt ggc ggt
tct ggc 672Leu Glu Ile Lys Arg Ala Asp Ala Ala Pro Gly Gly Gly Gly
Ser Gly 210 215 220 ggt ggc ggt tct gga ggc ggt ggc tct gag gtg cag
ctg gtg gag tct 720Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln
Leu Val Glu Ser 225 230 235 240 ggg gga ggc tta gtg aag cct gga ggg
tcc cgg aaa ctc tcc tgt gca 768Gly Gly Gly Leu Val Lys Pro Gly Gly
Ser Arg Lys Leu Ser Cys Ala 245 250 255 gcc tct gga ttc act ttc agt
aac tat gga atg cac tgg gtc cgt cag 816Ala Ser Gly Phe Thr Phe Ser
Asn Tyr Gly Met His Trp Val Arg Gln 260 265 270 gct cca gag aag ggg
ctg gag tgg gtt gca tac att agt agt ggc agt 864Ala Pro Glu Lys Gly
Leu Glu Trp Val Ala Tyr Ile Ser Ser Gly Ser 275 280 285 agt acc atc
tac tat gca gac aca gtg aag ggc cga ttc acc atc tcc 912Ser Thr Ile
Tyr Tyr Ala Asp Thr Val Lys Gly Arg Phe Thr Ile Ser 290 295 300 aga
gac aat gcc aag aac acc ctg ttc ctg caa atg acc agt cta agg 960Arg
Asp Asn Ala Lys Asn Thr Leu Phe Leu Gln Met Thr Ser Leu Arg 305 310
315 320 tct gag gac aca gcc atg cgg ggg tta cta ctt gac tac tgg ggc
caa 1008Ser Glu Asp Thr Ala Met Arg Gly Leu Leu Leu Asp Tyr Trp Gly
Gln 325 330 335 ggc acc act ctc aca gtc tcc tca cta gaa caa aaa ctc
atc tca gaa 1056Gly Thr Thr Leu Thr Val Ser Ser Leu Glu Gln Lys Leu
Ile Ser Glu 340 345 350 gag gat ctg aat agc gcc gtc gac gct tcc acc
aag ggc cca tcc gtc 1104Glu Asp Leu Asn Ser Ala Val Asp Ala Ser Thr
Lys Gly Pro Ser Val 355 360 365 ttc ccc ctg gcg ccc ctg gag tct tcc
gga tcc atg aag ctc tcc ctg 1152Phe Pro Leu Ala Pro Leu Glu Ser Ser
Gly Ser Met Lys Leu Ser Leu 370 375 380 gtg gcc gcg atg ctg ctg ctg
ctc agc gcg gcg cgg gcc gag gag gag 1200Val Ala Ala Met Leu Leu Leu
Leu Ser Ala Ala Arg Ala Glu Glu Glu 385 390 395 400 gac aag aag gag
gac gtg ggc acg gtg gtc ggc atc gac ctg ggg acc 1248Asp Lys Lys Glu
Asp Val Gly Thr Val Val Gly Ile Asp Leu Gly Thr 405 410 415 acc tac
tcc tgc gtc ggc gtg ttc aag aac ggc cgc gtg gag atc atc 1296Thr Tyr
Ser Cys Val Gly Val Phe Lys Asn Gly Arg Val Glu Ile Ile 420 425 430
gcc aac gat cag ggc aac cgc atc acg ccg tcc tat gtc gcc ttc act
1344Ala Asn Asp Gln Gly Asn Arg Ile Thr Pro Ser Tyr Val Ala Phe Thr
435 440 445 cct gaa ggg gaa cgt ctg att ggc gat gcc gcc aag aac cag
ctc acc 1392Pro Glu Gly Glu Arg Leu Ile Gly Asp Ala Ala Lys Asn Gln
Leu Thr 450 455 460 tcc aac ccc gag aac acg gtc ttt gac gcc aag cgg
ctc atc ggc cgc 1440Ser Asn Pro Glu Asn Thr Val Phe Asp Ala Lys Arg
Leu Ile Gly Arg 465 470 475 480 acg tgg aat gac ccg tct gtg cag cag
gac atc aag ttc ttg ccg ttc 1488Thr Trp Asn Asp Pro Ser Val Gln Gln
Asp Ile Lys Phe Leu Pro Phe 485 490 495 aag gtg gtt gaa aag aaa act
aaa cca tac att caa gtt gat att gga 1536Lys Val Val Glu Lys Lys Thr
Lys Pro Tyr Ile Gln Val Asp Ile Gly 500 505 510 ggt ggg caa aca aag
aca ttt gct cct gaa gaa att tct gcc atg gtt 1584Gly Gly Gln Thr Lys
Thr Phe Ala Pro Glu Glu Ile Ser Ala Met Val 515 520 525 ctc act aaa
atg aaa gaa acc gct gag gct tat ttg gga aag aag gtt 1632Leu Thr Lys
Met Lys Glu Thr Ala Glu Ala Tyr Leu Gly Lys Lys Val 530 535 540 acc
cat gca gtt gtt act gta cca gcc tat ttt aat gat gcc caa cgc 1680Thr
His Ala Val Val Thr Val Pro Ala Tyr Phe Asn Asp Ala Gln Arg 545 550
555 560 caa gca acc aaa gac gct gga act att gct ggc cta aat gtt atg
agg 1728Gln Ala Thr Lys Asp Ala Gly Thr Ile Ala Gly Leu Asn Val Met
Arg 565 570 575 atc atc aac gag cct acg gca gct gct att gct tat ggc
ctg gat aag 1776Ile Ile Asn Glu Pro Thr Ala Ala Ala Ile Ala Tyr Gly
Leu Asp Lys 580 585 590 agg gag ggg gag aag aac atc ctg gtg ttt gac
ctg ggt ggc gga acc 1824Arg Glu Gly Glu Lys Asn Ile Leu Val Phe Asp
Leu Gly Gly Gly Thr 595 600 605 ttc gat gtg tct ctt ctc acc att gac
aat ggt gtc ttc gaa gtt gtg 1872Phe Asp Val Ser Leu Leu Thr Ile Asp
Asn Gly Val Phe Glu Val Val 610 615 620 gcc act aat gga gat act cat
ctg ggt gga gaa gac ttt gac cag cgt 1920Ala Thr Asn Gly Asp Thr His
Leu Gly Gly Glu Asp Phe Asp Gln Arg 625 630 635 640 gtc atg gaa cac
ttc atc aaa ctg tac aaa aag aag acg ggc aaa gat 1968Val Met Glu His
Phe Ile Lys Leu Tyr Lys Lys Lys Thr Gly Lys Asp 645 650 655 gtc agg
aaa gac aat aga gct gtg cag aaa ctc cgg cgc gag gta gaa 2016Val Arg
Lys Asp Asn Arg Ala Val Gln Lys Leu Arg Arg Glu Val Glu 660 665 670
aag gcc aaa cgg gcc ctg tct tct cag cat caa gca aga att gaa att
2064Lys Ala Lys Arg Ala Leu Ser Ser Gln His Gln Ala Arg Ile Glu Ile
675 680 685 gag tcc ttc tat gaa gga gaa gac ttt tct gag acc ctg act
cgg gcc 2112Glu Ser Phe Tyr Glu Gly Glu Asp Phe Ser Glu Thr Leu Thr
Arg Ala 690 695 700 aaa ttt gaa gag ctc aac atg gat ctg ttc cgg tct
act atg aag ccc 2160Lys Phe Glu Glu Leu Asn Met Asp Leu Phe Arg Ser
Thr Met Lys Pro 705 710 715 720 gtc cag aaa gtg ttg gaa gat tct gat
ttg aag aag tct gat att gat 2208Val Gln Lys Val Leu Glu Asp Ser Asp
Leu Lys Lys Ser Asp Ile Asp 725 730 735 gaa att gtt ctt gtt ggt ggc
tcg act cga att cca aag att cag caa 2256Glu Ile Val Leu Val Gly Gly
Ser Thr Arg Ile Pro Lys Ile Gln Gln 740 745 750 ctg gtt aaa gag ttc
ttc aat ggc aag gaa cca tcc cgt ggc ata aac 2304Leu Val Lys Glu Phe
Phe Asn Gly Lys Glu Pro Ser Arg Gly Ile Asn 755 760 765 cca gat gaa
gct gta gcg tat ggt gct gct gtc cag gct ggt gtg ctc 2352Pro Asp Glu
Ala Val Ala Tyr Gly Ala Ala Val Gln Ala Gly Val Leu 770 775 780 tct
ggt gat caa gat aca ggt gac ctg gta ctg ctt gat gta tgt ccc 2400Ser
Gly Asp Gln Asp Thr Gly Asp Leu Val Leu Leu Asp Val Cys Pro 785 790
795 800 ctt aca ctt ggt att gaa act gtg gga ggt gtc atg acc aaa ctg
att 2448Leu Thr Leu Gly Ile Glu Thr Val Gly Gly Val Met Thr Lys Leu
Ile 805 810 815 cca agg aac aca gtg gtg cct acc aag aag tct cag atc
ttt tct aca 2496Pro Arg Asn Thr Val Val Pro Thr Lys Lys Ser Gln Ile
Phe Ser Thr 820 825 830 gct tct gat aat caa cca act gtt aca atc aag
gtc tat gaa ggt gaa 2544Ala Ser Asp Asn Gln Pro Thr Val Thr Ile Lys
Val Tyr Glu Gly Glu 835 840 845 aga ccc ctg aca aaa gac aat cat ctt
ctg ggt aca ttt gat ctg act 2592Arg Pro Leu Thr Lys Asp Asn His Leu
Leu Gly Thr Phe Asp Leu Thr 850 855 860 gga att cct cct gct cct cgt
ggg gtc cca cag att gaa gtc acc ttt 2640Gly Ile Pro Pro Ala Pro Arg
Gly Val Pro Gln Ile Glu Val Thr Phe 865 870 875 880 gag ata gat gtg
aat ggt att ctt cga gtg aca gct gaa gac aag ggt 2688Glu Ile Asp Val
Asn Gly Ile Leu Arg Val Thr Ala Glu Asp Lys Gly 885 890 895 aca ggg
aac aaa aat aag atc aca atc acc aat gac cag aat cgc ctg 2736Thr Gly
Asn Lys Asn Lys Ile Thr Ile Thr Asn Asp Gln Asn Arg Leu 900 905 910
aca cct gaa gaa atc gaa agg atg gtt aat gat gct gag aag ttt gct
2784Thr Pro Glu Glu Ile Glu Arg Met Val Asn Asp Ala Glu Lys Phe Ala
915 920 925 gag gaa gac aaa aag ctc aag gag cgc att gat act aga aat
gag ttg 2832Glu Glu Asp Lys Lys Leu Lys Glu Arg Ile Asp Thr Arg Asn
Glu Leu 930 935 940 gaa agc tat gcc tat tct cta aag aat cag att gga
gat aaa gaa aag 2880Glu Ser Tyr Ala Tyr Ser Leu Lys Asn Gln Ile Gly
Asp Lys Glu Lys 945 950 955 960 ctg gga ggt aaa ctt tcc tct gaa gat
aag gag acc atg gaa aaa gct 2928Leu Gly Gly Lys Leu Ser Ser Glu Asp
Lys Glu Thr Met Glu Lys Ala 965 970 975 gta gaa gaa aag att gaa tgg
ctg gaa agc cac caa gat gct gac att 2976Val Glu Glu Lys Ile Glu Trp
Leu Glu Ser His Gln Asp Ala Asp Ile 980 985 990 gaa gac ttc aaa gct
aag aag aag gaa ctg gaa gaa att gtt caa cca 3024Glu Asp Phe Lys Ala
Lys Lys Lys Glu Leu Glu Glu Ile Val Gln Pro 995 1000 1005 att atc
agc aaa ctc tat gga agt gca ggc cct ccc cca act ggt 3069Ile Ile Ser
Lys Leu Tyr Gly Ser Ala Gly Pro Pro Pro Thr Gly 1010 1015 1020 gaa
gag gat aca gca gaa aaa gat gag ttg tag tctaga 3108Glu Glu Asp Thr
Ala Glu Lys Asp Glu Leu 1025 1030 321033PRTArtificial
SequenceSynthetic Construct 32Met Arg Phe Pro Ser Leu Phe Thr Ala
Val Leu Phe Ala Ala Ser Ser 1 5 10 15 Ala Leu Ala Ala Pro Val Asn
Thr Thr Thr Glu Asp Glu Thr Ala Gln 20 25 30 Ile Pro Ala Glu Ala
Val Ile Gly Tyr Ser Asp Leu Glu Gly Asp Phe 35 40 45 Asp Val Ala
Val Leu Pro Phe Ser Asn Ser Thr Asn Asn Gly Leu Leu 50 55 60 Phe
Ile Asn Thr Thr Ile Ala Ser Ile Ala Ala Lys Glu Glu Gly Val 65 70
75 80 Ser Leu Glu Lys Arg Glu Ala Glu Ala Glu Phe His His His His
His 85 90 95 His Ala Gly Ile His Asp Ile Val Leu Thr Gln Ser Pro
Ala Ser Leu 100 105 110 Ala Val Ser Leu Gly Gln Arg Ala Thr Ile Ser
Cys Arg Ala Ser Lys 115 120 125 Ser Val Ser Thr Ser Ser Tyr Ser Tyr
Met His Trp Tyr Gln Gln Lys 130 135 140 Pro Gly Gln Pro Pro Lys Leu
Leu Ile Lys Tyr Ala Ser Tyr Leu Glu 145 150 155 160 Ser Gly Val Pro
Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe 165 170 175 Thr Leu
Asn Ile His Pro Val Glu Glu Glu Asp Ala Ala Thr Tyr Tyr 180 185 190
Cys Gln His Ser Arg Glu Phe Pro Trp Thr Phe Gly Gly Gly Thr Lys 195
200 205 Leu Glu Ile Lys Arg Ala Asp Ala Ala Pro Gly Gly Gly Gly Ser
Gly 210 215 220 Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu
Val Glu Ser 225 230 235 240 Gly Gly Gly Leu Val Lys Pro Gly Gly Ser
Arg Lys Leu Ser Cys Ala 245 250 255 Ala Ser Gly Phe Thr Phe Ser Asn
Tyr Gly Met His Trp Val Arg Gln 260 265 270 Ala Pro Glu Lys Gly Leu
Glu Trp Val Ala Tyr Ile Ser Ser Gly Ser 275 280 285 Ser Thr Ile Tyr
Tyr Ala Asp Thr Val Lys Gly Arg Phe Thr Ile Ser 290 295 300 Arg Asp
Asn Ala Lys Asn Thr Leu Phe Leu Gln Met Thr Ser Leu Arg 305 310 315
320 Ser Glu Asp Thr Ala Met Arg Gly Leu Leu Leu Asp Tyr Trp Gly Gln
325 330 335 Gly Thr Thr Leu Thr Val Ser Ser Leu Glu Gln Lys Leu Ile
Ser Glu 340 345 350 Glu Asp Leu Asn Ser Ala Val Asp Ala Ser Thr Lys
Gly Pro Ser Val 355 360
365 Phe Pro Leu Ala Pro Leu Glu Ser Ser Gly Ser Met Lys Leu Ser Leu
370 375 380 Val Ala Ala Met Leu Leu Leu Leu Ser Ala Ala Arg Ala Glu
Glu Glu 385 390 395 400 Asp Lys Lys Glu Asp Val Gly Thr Val Val Gly
Ile Asp Leu Gly Thr 405 410 415 Thr Tyr Ser Cys Val Gly Val Phe Lys
Asn Gly Arg Val Glu Ile Ile 420 425 430 Ala Asn Asp Gln Gly Asn Arg
Ile Thr Pro Ser Tyr Val Ala Phe Thr 435 440 445 Pro Glu Gly Glu Arg
Leu Ile Gly Asp Ala Ala Lys Asn Gln Leu Thr 450 455 460 Ser Asn Pro
Glu Asn Thr Val Phe Asp Ala Lys Arg Leu Ile Gly Arg 465 470 475 480
Thr Trp Asn Asp Pro Ser Val Gln Gln Asp Ile Lys Phe Leu Pro Phe 485
490 495 Lys Val Val Glu Lys Lys Thr Lys Pro Tyr Ile Gln Val Asp Ile
Gly 500 505 510 Gly Gly Gln Thr Lys Thr Phe Ala Pro Glu Glu Ile Ser
Ala Met Val 515 520 525 Leu Thr Lys Met Lys Glu Thr Ala Glu Ala Tyr
Leu Gly Lys Lys Val 530 535 540 Thr His Ala Val Val Thr Val Pro Ala
Tyr Phe Asn Asp Ala Gln Arg 545 550 555 560 Gln Ala Thr Lys Asp Ala
Gly Thr Ile Ala Gly Leu Asn Val Met Arg 565 570 575 Ile Ile Asn Glu
Pro Thr Ala Ala Ala Ile Ala Tyr Gly Leu Asp Lys 580 585 590 Arg Glu
Gly Glu Lys Asn Ile Leu Val Phe Asp Leu Gly Gly Gly Thr 595 600 605
Phe Asp Val Ser Leu Leu Thr Ile Asp Asn Gly Val Phe Glu Val Val 610
615 620 Ala Thr Asn Gly Asp Thr His Leu Gly Gly Glu Asp Phe Asp Gln
Arg 625 630 635 640 Val Met Glu His Phe Ile Lys Leu Tyr Lys Lys Lys
Thr Gly Lys Asp 645 650 655 Val Arg Lys Asp Asn Arg Ala Val Gln Lys
Leu Arg Arg Glu Val Glu 660 665 670 Lys Ala Lys Arg Ala Leu Ser Ser
Gln His Gln Ala Arg Ile Glu Ile 675 680 685 Glu Ser Phe Tyr Glu Gly
Glu Asp Phe Ser Glu Thr Leu Thr Arg Ala 690 695 700 Lys Phe Glu Glu
Leu Asn Met Asp Leu Phe Arg Ser Thr Met Lys Pro 705 710 715 720 Val
Gln Lys Val Leu Glu Asp Ser Asp Leu Lys Lys Ser Asp Ile Asp 725 730
735 Glu Ile Val Leu Val Gly Gly Ser Thr Arg Ile Pro Lys Ile Gln Gln
740 745 750 Leu Val Lys Glu Phe Phe Asn Gly Lys Glu Pro Ser Arg Gly
Ile Asn 755 760 765 Pro Asp Glu Ala Val Ala Tyr Gly Ala Ala Val Gln
Ala Gly Val Leu 770 775 780 Ser Gly Asp Gln Asp Thr Gly Asp Leu Val
Leu Leu Asp Val Cys Pro 785 790 795 800 Leu Thr Leu Gly Ile Glu Thr
Val Gly Gly Val Met Thr Lys Leu Ile 805 810 815 Pro Arg Asn Thr Val
Val Pro Thr Lys Lys Ser Gln Ile Phe Ser Thr 820 825 830 Ala Ser Asp
Asn Gln Pro Thr Val Thr Ile Lys Val Tyr Glu Gly Glu 835 840 845 Arg
Pro Leu Thr Lys Asp Asn His Leu Leu Gly Thr Phe Asp Leu Thr 850 855
860 Gly Ile Pro Pro Ala Pro Arg Gly Val Pro Gln Ile Glu Val Thr Phe
865 870 875 880 Glu Ile Asp Val Asn Gly Ile Leu Arg Val Thr Ala Glu
Asp Lys Gly 885 890 895 Thr Gly Asn Lys Asn Lys Ile Thr Ile Thr Asn
Asp Gln Asn Arg Leu 900 905 910 Thr Pro Glu Glu Ile Glu Arg Met Val
Asn Asp Ala Glu Lys Phe Ala 915 920 925 Glu Glu Asp Lys Lys Leu Lys
Glu Arg Ile Asp Thr Arg Asn Glu Leu 930 935 940 Glu Ser Tyr Ala Tyr
Ser Leu Lys Asn Gln Ile Gly Asp Lys Glu Lys 945 950 955 960 Leu Gly
Gly Lys Leu Ser Ser Glu Asp Lys Glu Thr Met Glu Lys Ala 965 970 975
Val Glu Glu Lys Ile Glu Trp Leu Glu Ser His Gln Asp Ala Asp Ile 980
985 990 Glu Asp Phe Lys Ala Lys Lys Lys Glu Leu Glu Glu Ile Val Gln
Pro 995 1000 1005 Ile Ile Ser Lys Leu Tyr Gly Ser Ala Gly Pro Pro
Pro Thr Gly 1010 1015 1020 Glu Glu Asp Thr Ala Glu Lys Asp Glu Leu
1025 1030 331033PRTArtificial Sequence3E10 Fv-GRP78 amino acid
sequence for a chimeric protein comprising a single chain Fv
antibody fragment derived from Mus musculus 3E10 hybridoma and
GRP78 from Homo sapiens joined by human CH1 linker and swivel
sequence 33Met Arg Phe Pro Ser Leu Phe Thr Ala Val Leu Phe Ala Ala
Ser Ser 1 5 10 15 Ala Leu Ala Ala Pro Val Asn Thr Thr Thr Glu Asp
Glu Thr Ala Gln 20 25 30 Ile Pro Ala Glu Ala Val Ile Gly Tyr Ser
Asp Leu Glu Gly Asp Phe 35 40 45 Asp Val Ala Val Leu Pro Phe Ser
Asn Ser Thr Asn Asn Gly Leu Leu 50 55 60 Phe Ile Asn Thr Thr Ile
Ala Ser Ile Ala Ala Lys Glu Glu Gly Val 65 70 75 80 Ser Leu Glu Lys
Arg Glu Ala Glu Ala Glu Phe His His His His His 85 90 95 His Ala
Gly Ile His Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu 100 105 110
Ala Val Ser Leu Gly Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Lys 115
120 125 Ser Val Ser Thr Ser Ser Tyr Ser Tyr Met His Trp Tyr Gln Gln
Lys 130 135 140 Pro Gly Gln Pro Pro Lys Leu Leu Ile Lys Tyr Ala Ser
Tyr Leu Glu 145 150 155 160 Ser Gly Val Pro Ala Arg Phe Ser Gly Ser
Gly Ser Gly Thr Asp Phe 165 170 175 Thr Leu Asn Ile His Pro Val Glu
Glu Glu Asp Ala Ala Thr Tyr Tyr 180 185 190 Cys Gln His Ser Arg Glu
Phe Pro Trp Thr Phe Gly Gly Gly Thr Lys 195 200 205 Leu Glu Ile Lys
Arg Ala Asp Ala Ala Pro Gly Gly Gly Gly Ser Gly 210 215 220 Gly Gly
Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser 225 230 235
240 Gly Gly Gly Leu Val Lys Pro Gly Gly Ser Arg Lys Leu Ser Cys Ala
245 250 255 Ala Ser Gly Phe Thr Phe Ser Asn Tyr Gly Met His Trp Val
Arg Gln 260 265 270 Ala Pro Glu Lys Gly Leu Glu Trp Val Ala Tyr Ile
Ser Ser Gly Ser 275 280 285 Ser Thr Ile Tyr Tyr Ala Asp Thr Val Lys
Gly Arg Phe Thr Ile Ser 290 295 300 Arg Asp Asn Ala Lys Asn Thr Leu
Phe Leu Gln Met Thr Ser Leu Arg 305 310 315 320 Ser Glu Asp Thr Ala
Met Arg Gly Leu Leu Leu Asp Tyr Trp Gly Gln 325 330 335 Gly Thr Thr
Leu Thr Val Ser Ser Leu Glu Gln Lys Leu Ile Ser Glu 340 345 350 Glu
Asp Leu Asn Ser Ala Val Asp Ala Ser Thr Lys Gly Pro Ser Val 355 360
365 Phe Pro Leu Ala Pro Leu Glu Ser Ser Gly Ser Met Lys Leu Ser Leu
370 375 380 Val Ala Ala Met Leu Leu Leu Leu Ser Ala Ala Arg Ala Glu
Glu Glu 385 390 395 400 Asp Lys Lys Glu Asp Val Gly Thr Val Val Gly
Ile Asp Leu Gly Thr 405 410 415 Thr Tyr Ser Cys Val Gly Val Phe Lys
Asn Gly Arg Val Glu Ile Ile 420 425 430 Ala Asn Asp Gln Gly Asn Arg
Ile Thr Pro Ser Tyr Val Ala Phe Thr 435 440 445 Pro Glu Gly Glu Arg
Leu Ile Gly Asp Ala Ala Lys Asn Gln Leu Thr 450 455 460 Ser Asn Pro
Glu Asn Thr Val Phe Asp Ala Lys Arg Leu Ile Gly Arg 465 470 475 480
Thr Trp Asn Asp Pro Ser Val Gln Gln Asp Ile Lys Phe Leu Pro Phe 485
490 495 Lys Val Val Glu Lys Lys Thr Lys Pro Tyr Ile Gln Val Asp Ile
Gly 500 505 510 Gly Gly Gln Thr Lys Thr Phe Ala Pro Glu Glu Ile Ser
Ala Met Val 515 520 525 Leu Thr Lys Met Lys Glu Thr Ala Glu Ala Tyr
Leu Gly Lys Lys Val 530 535 540 Thr His Ala Val Val Thr Val Pro Ala
Tyr Phe Asn Asp Ala Gln Arg 545 550 555 560 Gln Ala Thr Lys Asp Ala
Gly Thr Ile Ala Gly Leu Asn Val Met Arg 565 570 575 Ile Ile Asn Glu
Pro Thr Ala Ala Ala Ile Ala Tyr Gly Leu Asp Lys 580 585 590 Arg Glu
Gly Glu Lys Asn Ile Leu Val Phe Asp Leu Gly Gly Gly Thr 595 600 605
Phe Asp Val Ser Leu Leu Thr Ile Asp Asn Gly Val Phe Glu Val Val 610
615 620 Ala Thr Asn Gly Asp Thr His Leu Gly Gly Glu Asp Phe Asp Gln
Arg 625 630 635 640 Val Met Glu His Phe Ile Lys Leu Tyr Lys Lys Lys
Thr Gly Lys Asp 645 650 655 Val Arg Lys Asp Asn Arg Ala Val Gln Lys
Leu Arg Arg Glu Val Glu 660 665 670 Lys Ala Lys Arg Ala Leu Ser Ser
Gln His Gln Ala Arg Ile Glu Ile 675 680 685 Glu Ser Phe Tyr Glu Gly
Glu Asp Phe Ser Glu Thr Leu Thr Arg Ala 690 695 700 Lys Phe Glu Glu
Leu Asn Met Asp Leu Phe Arg Ser Thr Met Lys Pro 705 710 715 720 Val
Gln Lys Val Leu Glu Asp Ser Asp Leu Lys Lys Ser Asp Ile Asp 725 730
735 Glu Ile Val Leu Val Gly Gly Ser Thr Arg Ile Pro Lys Ile Gln Gln
740 745 750 Leu Val Lys Glu Phe Phe Asn Gly Lys Glu Pro Ser Arg Gly
Ile Asn 755 760 765 Pro Asp Glu Ala Val Ala Tyr Gly Ala Ala Val Gln
Ala Gly Val Leu 770 775 780 Ser Gly Asp Gln Asp Thr Gly Asp Leu Val
Leu Leu Asp Val Cys Pro 785 790 795 800 Leu Thr Leu Gly Ile Glu Thr
Val Gly Gly Val Met Thr Lys Leu Ile 805 810 815 Pro Arg Asn Thr Val
Val Pro Thr Lys Lys Ser Gln Ile Phe Ser Thr 820 825 830 Ala Ser Asp
Asn Gln Pro Thr Val Thr Ile Lys Val Tyr Glu Gly Glu 835 840 845 Arg
Pro Leu Thr Lys Asp Asn His Leu Leu Gly Thr Phe Asp Leu Thr 850 855
860 Gly Ile Pro Pro Ala Pro Arg Gly Val Pro Gln Ile Glu Val Thr Phe
865 870 875 880 Glu Ile Asp Val Asn Gly Ile Leu Arg Val Thr Ala Glu
Asp Lys Gly 885 890 895 Thr Gly Asn Lys Asn Lys Ile Thr Ile Thr Asn
Asp Gln Asn Arg Leu 900 905 910 Thr Pro Glu Glu Ile Glu Arg Met Val
Asn Asp Ala Glu Lys Phe Ala 915 920 925 Glu Glu Asp Lys Lys Leu Lys
Glu Arg Ile Asp Thr Arg Asn Glu Leu 930 935 940 Glu Ser Tyr Ala Tyr
Ser Leu Lys Asn Gln Ile Gly Asp Lys Glu Lys 945 950 955 960 Leu Gly
Gly Lys Leu Ser Ser Glu Asp Lys Glu Thr Met Glu Lys Ala 965 970 975
Val Glu Glu Lys Ile Glu Trp Leu Glu Ser His Gln Asp Ala Asp Ile 980
985 990 Glu Asp Phe Lys Ala Lys Lys Lys Glu Leu Glu Glu Ile Val Gln
Pro 995 1000 1005 Ile Ile Ser Lys Leu Tyr Gly Ser Ala Gly Pro Pro
Pro Thr Gly 1010 1015 1020 Glu Glu Asp Thr Ala Glu Lys Asp Glu Leu
1025 1030 343342DNAArtificial Sequence3E10 Fv-Hsp90 nucleotide
sequence encoding a chimeric protein comprising a single chain Fv
antibody fragment derived from Mus musculus 3E10 hybridoma and
Hsp90 from Homo sapiens joined by human CH1 linker and swivel
sequence 34atg aga ttt cct tca ctt ttt act gct gtt tta ttc gca gca
tcc tcc 48Met Arg Phe Pro Ser Leu Phe Thr Ala Val Leu Phe Ala Ala
Ser Ser 1 5 10 15 gca tta gct gct cca gtc aac act aca aca gaa gat
gaa acg gca caa 96Ala Leu Ala Ala Pro Val Asn Thr Thr Thr Glu Asp
Glu Thr Ala Gln 20 25 30 att ccg gct gaa gct gtc atc ggt tac tca
gat tta gaa ggg gat ttc 144Ile Pro Ala Glu Ala Val Ile Gly Tyr Ser
Asp Leu Glu Gly Asp Phe 35 40 45 gat gtt gct gtt ttg cca ttt tcc
aac agc aca aat aac ggg tta ttg 192Asp Val Ala Val Leu Pro Phe Ser
Asn Ser Thr Asn Asn Gly Leu Leu 50 55 60 ttt ata aat act act att
gcc agc att gct gct aaa gaa gaa ggg gta 240Phe Ile Asn Thr Thr Ile
Ala Ser Ile Ala Ala Lys Glu Glu Gly Val 65 70 75 80 tct ctc gag aaa
aga gag gct gaa gct gaa ttc cat cac cat cac cat 288Ser Leu Glu Lys
Arg Glu Ala Glu Ala Glu Phe His His His His His 85 90 95 cac gca
ggg att cac gac att gtc ctg aca cag tct cct gct tcc tta 336His Ala
Gly Ile His Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu 100 105 110
gct gta tct ctg ggg cag agg gcc acc atc tcc tgc agg gcc agc aaa
384Ala Val Ser Leu Gly Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Lys
115 120 125 agt gtc agt aca tct agc tat agt tac atg cac tgg tac caa
cag aaa 432Ser Val Ser Thr Ser Ser Tyr Ser Tyr Met His Trp Tyr Gln
Gln Lys 130 135 140 cca gga cag cca ccc aaa ctc ctc atc aag tat gca
tcc tac cta gaa 480Pro Gly Gln Pro Pro Lys Leu Leu Ile Lys Tyr Ala
Ser Tyr Leu Glu 145 150 155 160 tct ggg gtt cct gcc agg ttc agt ggc
agt ggg tct ggg aca gac ttc 528Ser Gly Val Pro Ala Arg Phe Ser Gly
Ser Gly Ser Gly Thr Asp Phe 165 170 175 acc ctc aac atc cat cct gtg
gag gag gag gat gct gca aca tat tac 576Thr Leu Asn Ile His Pro Val
Glu Glu Glu Asp Ala Ala Thr Tyr Tyr 180 185 190 tgt cag cac agt agg
gag ttt ccg tgg acg ttc ggt gga ggc acc aag 624Cys Gln His Ser Arg
Glu Phe Pro Trp Thr Phe Gly Gly Gly Thr Lys 195 200 205 ctg gaa atc
aaa cgg gct gat gct gca ccc ggg ggt ggc ggt tct ggc 672Leu Glu Ile
Lys Arg Ala Asp Ala Ala Pro Gly Gly Gly Gly Ser Gly 210 215 220 ggt
ggc ggt tct gga ggc ggt ggc tct gag gtg cag ctg gtg gag tct 720Gly
Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser 225 230
235 240 ggg gga ggc tta gtg aag cct gga ggg tcc cgg aaa ctc tcc tgt
gca 768Gly Gly Gly Leu Val Lys Pro Gly Gly Ser Arg Lys Leu Ser Cys
Ala 245 250 255 gcc tct gga ttc act ttc agt aac tat gga atg cac tgg
gtc cgt cag 816Ala Ser Gly Phe Thr Phe Ser Asn Tyr Gly Met His Trp
Val Arg Gln 260 265 270 gct cca gag aag ggg ctg gag tgg gtt gca tac
att agt agt ggc agt 864Ala Pro Glu Lys Gly Leu Glu Trp Val Ala Tyr
Ile Ser Ser Gly Ser 275 280 285 agt acc atc tac tat gca gac aca gtg
aag ggc cga ttc acc atc tcc 912Ser Thr Ile Tyr Tyr Ala Asp Thr Val
Lys Gly Arg Phe Thr Ile Ser 290 295 300 aga gac aat gcc aag aac acc
ctg ttc ctg caa atg acc agt cta agg
960Arg Asp Asn Ala Lys Asn Thr Leu Phe Leu Gln Met Thr Ser Leu Arg
305 310 315 320 tct gag gac aca gcc atg cgg ggg tta cta ctt gac tac
tgg ggc caa 1008Ser Glu Asp Thr Ala Met Arg Gly Leu Leu Leu Asp Tyr
Trp Gly Gln 325 330 335 ggc acc act ctc aca gtc tcc tca cta gaa caa
aaa ctc atc tca gaa 1056Gly Thr Thr Leu Thr Val Ser Ser Leu Glu Gln
Lys Leu Ile Ser Glu 340 345 350 gag gat ctg aat agc gcc gtc gac gct
tcc acc aag ggc cca tcc gtc 1104Glu Asp Leu Asn Ser Ala Val Asp Ala
Ser Thr Lys Gly Pro Ser Val 355 360 365 ttc ccc ctg gcg ccc ctg gag
tct tcc gga tcc atg cct gag gaa acc 1152Phe Pro Leu Ala Pro Leu Glu
Ser Ser Gly Ser Met Pro Glu Glu Thr 370 375 380 cag acc caa gac caa
ccg atg gag gag gag gag gtt gag acg ttc gcc 1200Gln Thr Gln Asp Gln
Pro Met Glu Glu Glu Glu Val Glu Thr Phe Ala 385 390 395 400 ttt cag
gca gaa att gcc cag ttg atg tca ttg atc atc aat act ttc 1248Phe Gln
Ala Glu Ile Ala Gln Leu Met Ser Leu Ile Ile Asn Thr Phe 405 410 415
tac tcg aac aaa gag atc ttt ctg aga gag ctc att tca aat tca tca
1296Tyr Ser Asn Lys Glu Ile Phe Leu Arg Glu Leu Ile Ser Asn Ser Ser
420 425 430 gat gca ttg gac aaa atc cgg tat gaa agc ttg aca gat ccc
agt aaa 1344Asp Ala Leu Asp Lys Ile Arg Tyr Glu Ser Leu Thr Asp Pro
Ser Lys 435 440 445 tta gac tct ggg aaa gag ctg cat att aac ctt ata
ccg aac aaa caa 1392Leu Asp Ser Gly Lys Glu Leu His Ile Asn Leu Ile
Pro Asn Lys Gln 450 455 460 gat cga act ctc act att gtg gat act gga
att gga atg acc aag gct 1440Asp Arg Thr Leu Thr Ile Val Asp Thr Gly
Ile Gly Met Thr Lys Ala 465 470 475 480 gac ttg atc aat aac ctt ggt
act atc gcc aag tct ggg acc aaa gcg 1488Asp Leu Ile Asn Asn Leu Gly
Thr Ile Ala Lys Ser Gly Thr Lys Ala 485 490 495 ttc atg gaa gct ttg
cag gct ggt gca gat atc tct atg att ggc cag 1536Phe Met Glu Ala Leu
Gln Ala Gly Ala Asp Ile Ser Met Ile Gly Gln 500 505 510 ttc ggt gtt
ggt ttt tat tct gct tat ttg gtt gct gag aaa gta act 1584Phe Gly Val
Gly Phe Tyr Ser Ala Tyr Leu Val Ala Glu Lys Val Thr 515 520 525 gtg
atc acc aaa cat aac gat gat gag cag tac gct tgg gag tcc tca 1632Val
Ile Thr Lys His Asn Asp Asp Glu Gln Tyr Ala Trp Glu Ser Ser 530 535
540 gca ggg gga tca ttc aca gtg agg aca gac aca ggt gaa cct atg ggt
1680Ala Gly Gly Ser Phe Thr Val Arg Thr Asp Thr Gly Glu Pro Met Gly
545 550 555 560 cgt gga aca aaa gtt atc cta cac ctg aaa gaa gac caa
act gag tac 1728Arg Gly Thr Lys Val Ile Leu His Leu Lys Glu Asp Gln
Thr Glu Tyr 565 570 575 ttg gag gaa cga aga ata aag gag att gtg aag
aaa cat tct cag ttt 1776Leu Glu Glu Arg Arg Ile Lys Glu Ile Val Lys
Lys His Ser Gln Phe 580 585 590 att gga tat ccc att act ctt ttt gtg
gag aag gaa cgt gat aaa gaa 1824Ile Gly Tyr Pro Ile Thr Leu Phe Val
Glu Lys Glu Arg Asp Lys Glu 595 600 605 gta agc gat gat gag gct gaa
gaa aag gaa gac aaa gaa gaa gaa aaa 1872Val Ser Asp Asp Glu Ala Glu
Glu Lys Glu Asp Lys Glu Glu Glu Lys 610 615 620 gaa aaa gaa gag aaa
gag tcg gaa gac aaa cct gaa att gaa gat gtt 1920Glu Lys Glu Glu Lys
Glu Ser Glu Asp Lys Pro Glu Ile Glu Asp Val 625 630 635 640 ggt tct
gat gag gaa gaa gaa aag aag gat ggt gac aag aag aag aag 1968Gly Ser
Asp Glu Glu Glu Glu Lys Lys Asp Gly Asp Lys Lys Lys Lys 645 650 655
aag aag att aag gaa aag tac atc gat caa gaa gag ctc aac aaa aca
2016Lys Lys Ile Lys Glu Lys Tyr Ile Asp Gln Glu Glu Leu Asn Lys Thr
660 665 670 aag ccc atc tgg acc aga aat ccc gac gat att act aat gag
gag tac 2064Lys Pro Ile Trp Thr Arg Asn Pro Asp Asp Ile Thr Asn Glu
Glu Tyr 675 680 685 gga gaa ttc tat aag agc ttg acc aat gac tgg gaa
gat cac ttg gca 2112Gly Glu Phe Tyr Lys Ser Leu Thr Asn Asp Trp Glu
Asp His Leu Ala 690 695 700 gtg aag cat ttt tca gtt gaa gga cag ttg
gaa ttc aga gcc ctt cta 2160Val Lys His Phe Ser Val Glu Gly Gln Leu
Glu Phe Arg Ala Leu Leu 705 710 715 720 ttt gtc cca cga cgt gct cct
ttt gat ctg ttt gaa aac aga aag aaa 2208Phe Val Pro Arg Arg Ala Pro
Phe Asp Leu Phe Glu Asn Arg Lys Lys 725 730 735 aag aac aac atc aaa
ttg tat gta cgc aga gtt ttc atc atg gat aac 2256Lys Asn Asn Ile Lys
Leu Tyr Val Arg Arg Val Phe Ile Met Asp Asn 740 745 750 tgt gag gag
cta atc cct gaa tat ctg aac ttc att aga ggg gtg gta 2304Cys Glu Glu
Leu Ile Pro Glu Tyr Leu Asn Phe Ile Arg Gly Val Val 755 760 765 gac
tcg gag gat ctc cct cta aac ata tcc cgt gag atg ttg caa caa 2352Asp
Ser Glu Asp Leu Pro Leu Asn Ile Ser Arg Glu Met Leu Gln Gln 770 775
780 agc aaa att ttg aaa gtt atc agg aag aat ttg gtc aaa aaa tgc tta
2400Ser Lys Ile Leu Lys Val Ile Arg Lys Asn Leu Val Lys Lys Cys Leu
785 790 795 800 gaa ctc ttt act gaa ctg gcg gaa gat aaa gag aac tac
aag aaa ttc 2448Glu Leu Phe Thr Glu Leu Ala Glu Asp Lys Glu Asn Tyr
Lys Lys Phe 805 810 815 tat gag cag ttc tct aaa aac ata aag ctt gga
ata cac gaa gac tct 2496Tyr Glu Gln Phe Ser Lys Asn Ile Lys Leu Gly
Ile His Glu Asp Ser 820 825 830 caa aat cgg aag aag ctt tca gag ctg
tta agg tac tac aca tct gcc 2544Gln Asn Arg Lys Lys Leu Ser Glu Leu
Leu Arg Tyr Tyr Thr Ser Ala 835 840 845 tct ggt gat gag atg gtt tct
ctc aag gac tac tgc acc aga atg aag 2592Ser Gly Asp Glu Met Val Ser
Leu Lys Asp Tyr Cys Thr Arg Met Lys 850 855 860 gag aac cag aaa cat
atc tat tat atc aca ggt gag acc aag gac cag 2640Glu Asn Gln Lys His
Ile Tyr Tyr Ile Thr Gly Glu Thr Lys Asp Gln 865 870 875 880 gta gct
aac tca gcc ttt gtg gaa cgt ctt cgg aaa cat ggc tta gaa 2688Val Ala
Asn Ser Ala Phe Val Glu Arg Leu Arg Lys His Gly Leu Glu 885 890 895
gtg atc tat atg att gag ccc att gat gag tac tgt gtc caa cag ctg
2736Val Ile Tyr Met Ile Glu Pro Ile Asp Glu Tyr Cys Val Gln Gln Leu
900 905 910 aag gaa ttt gag ggg aag act tta gtg tca gtc acc aaa gaa
ggc ctg 2784Lys Glu Phe Glu Gly Lys Thr Leu Val Ser Val Thr Lys Glu
Gly Leu 915 920 925 gaa ctt cca gag gat gaa gaa gag aaa aag aag cag
gaa gag aaa aaa 2832Glu Leu Pro Glu Asp Glu Glu Glu Lys Lys Lys Gln
Glu Glu Lys Lys 930 935 940 aca aag ttt gag aac ctc tgc aaa atc atg
aaa gac ata ttg gag aaa 2880Thr Lys Phe Glu Asn Leu Cys Lys Ile Met
Lys Asp Ile Leu Glu Lys 945 950 955 960 aaa gtt gaa aag gtg gtt gtg
tca aac cga ttg gtg aca tct cca tgc 2928Lys Val Glu Lys Val Val Val
Ser Asn Arg Leu Val Thr Ser Pro Cys 965 970 975 tgt att gtc aca agc
aca tat ggc tgg aca gca aac atg gag aga atc 2976Cys Ile Val Thr Ser
Thr Tyr Gly Trp Thr Ala Asn Met Glu Arg Ile 980 985 990 atg aaa gct
caa gcc cta aga gac aac tca aca atg ggt tac atg gca 3024Met Lys Ala
Gln Ala Leu Arg Asp Asn Ser Thr Met Gly Tyr Met Ala 995 1000 1005
gca aag aaa cac ctg gag ata aac cct gac cat tcc att att gag 3069Ala
Lys Lys His Leu Glu Ile Asn Pro Asp His Ser Ile Ile Glu 1010 1015
1020 acc tta agg caa aag gca gag gct gat aag aac gac aag tct gtg
3114Thr Leu Arg Gln Lys Ala Glu Ala Asp Lys Asn Asp Lys Ser Val
1025 1030 1035 aag gat ctg gtc atc ttg ctt tat gaa act gcg ctc ctg
tct tct 3159Lys Asp Leu Val Ile Leu Leu Tyr Glu Thr Ala Leu Leu Ser
Ser 1040 1045 1050 ggc ttc agt ctg gaa gat ccc cag aca cat gct aac
agg atc tac 3204Gly Phe Ser Leu Glu Asp Pro Gln Thr His Ala Asn Arg
Ile Tyr 1055 1060 1065 agg atg atc aaa ctt ggt ctg ggt att gat gaa
gat gac cct act 3249Arg Met Ile Lys Leu Gly Leu Gly Ile Asp Glu Asp
Asp Pro Thr 1070 1075 1080 gct gat gat acc agt gct gct gta act gaa
gaa atg cca ccc ctt 3294Ala Asp Asp Thr Ser Ala Ala Val Thr Glu Glu
Met Pro Pro Leu 1085 1090 1095 gaa gga gat gac gac aca tca cgc atg
gaa gaa gta gac taa tctaga 3342Glu Gly Asp Asp Asp Thr Ser Arg Met
Glu Glu Val Asp 1100 1105 1110 351111PRTArtificial
SequenceSynthetic Construct 35Met Arg Phe Pro Ser Leu Phe Thr Ala
Val Leu Phe Ala Ala Ser Ser 1 5 10 15 Ala Leu Ala Ala Pro Val Asn
Thr Thr Thr Glu Asp Glu Thr Ala Gln 20 25 30 Ile Pro Ala Glu Ala
Val Ile Gly Tyr Ser Asp Leu Glu Gly Asp Phe 35 40 45 Asp Val Ala
Val Leu Pro Phe Ser Asn Ser Thr Asn Asn Gly Leu Leu 50 55 60 Phe
Ile Asn Thr Thr Ile Ala Ser Ile Ala Ala Lys Glu Glu Gly Val 65 70
75 80 Ser Leu Glu Lys Arg Glu Ala Glu Ala Glu Phe His His His His
His 85 90 95 His Ala Gly Ile His Asp Ile Val Leu Thr Gln Ser Pro
Ala Ser Leu 100 105 110 Ala Val Ser Leu Gly Gln Arg Ala Thr Ile Ser
Cys Arg Ala Ser Lys 115 120 125 Ser Val Ser Thr Ser Ser Tyr Ser Tyr
Met His Trp Tyr Gln Gln Lys 130 135 140 Pro Gly Gln Pro Pro Lys Leu
Leu Ile Lys Tyr Ala Ser Tyr Leu Glu 145 150 155 160 Ser Gly Val Pro
Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe 165 170 175 Thr Leu
Asn Ile His Pro Val Glu Glu Glu Asp Ala Ala Thr Tyr Tyr 180 185 190
Cys Gln His Ser Arg Glu Phe Pro Trp Thr Phe Gly Gly Gly Thr Lys 195
200 205 Leu Glu Ile Lys Arg Ala Asp Ala Ala Pro Gly Gly Gly Gly Ser
Gly 210 215 220 Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu
Val Glu Ser 225 230 235 240 Gly Gly Gly Leu Val Lys Pro Gly Gly Ser
Arg Lys Leu Ser Cys Ala 245 250 255 Ala Ser Gly Phe Thr Phe Ser Asn
Tyr Gly Met His Trp Val Arg Gln 260 265 270 Ala Pro Glu Lys Gly Leu
Glu Trp Val Ala Tyr Ile Ser Ser Gly Ser 275 280 285 Ser Thr Ile Tyr
Tyr Ala Asp Thr Val Lys Gly Arg Phe Thr Ile Ser 290 295 300 Arg Asp
Asn Ala Lys Asn Thr Leu Phe Leu Gln Met Thr Ser Leu Arg 305 310 315
320 Ser Glu Asp Thr Ala Met Arg Gly Leu Leu Leu Asp Tyr Trp Gly Gln
325 330 335 Gly Thr Thr Leu Thr Val Ser Ser Leu Glu Gln Lys Leu Ile
Ser Glu 340 345 350 Glu Asp Leu Asn Ser Ala Val Asp Ala Ser Thr Lys
Gly Pro Ser Val 355 360 365 Phe Pro Leu Ala Pro Leu Glu Ser Ser Gly
Ser Met Pro Glu Glu Thr 370 375 380 Gln Thr Gln Asp Gln Pro Met Glu
Glu Glu Glu Val Glu Thr Phe Ala 385 390 395 400 Phe Gln Ala Glu Ile
Ala Gln Leu Met Ser Leu Ile Ile Asn Thr Phe 405 410 415 Tyr Ser Asn
Lys Glu Ile Phe Leu Arg Glu Leu Ile Ser Asn Ser Ser 420 425 430 Asp
Ala Leu Asp Lys Ile Arg Tyr Glu Ser Leu Thr Asp Pro Ser Lys 435 440
445 Leu Asp Ser Gly Lys Glu Leu His Ile Asn Leu Ile Pro Asn Lys Gln
450 455 460 Asp Arg Thr Leu Thr Ile Val Asp Thr Gly Ile Gly Met Thr
Lys Ala 465 470 475 480 Asp Leu Ile Asn Asn Leu Gly Thr Ile Ala Lys
Ser Gly Thr Lys Ala 485 490 495 Phe Met Glu Ala Leu Gln Ala Gly Ala
Asp Ile Ser Met Ile Gly Gln 500 505 510 Phe Gly Val Gly Phe Tyr Ser
Ala Tyr Leu Val Ala Glu Lys Val Thr 515 520 525 Val Ile Thr Lys His
Asn Asp Asp Glu Gln Tyr Ala Trp Glu Ser Ser 530 535 540 Ala Gly Gly
Ser Phe Thr Val Arg Thr Asp Thr Gly Glu Pro Met Gly 545 550 555 560
Arg Gly Thr Lys Val Ile Leu His Leu Lys Glu Asp Gln Thr Glu Tyr 565
570 575 Leu Glu Glu Arg Arg Ile Lys Glu Ile Val Lys Lys His Ser Gln
Phe 580 585 590 Ile Gly Tyr Pro Ile Thr Leu Phe Val Glu Lys Glu Arg
Asp Lys Glu 595 600 605 Val Ser Asp Asp Glu Ala Glu Glu Lys Glu Asp
Lys Glu Glu Glu Lys 610 615 620 Glu Lys Glu Glu Lys Glu Ser Glu Asp
Lys Pro Glu Ile Glu Asp Val 625 630 635 640 Gly Ser Asp Glu Glu Glu
Glu Lys Lys Asp Gly Asp Lys Lys Lys Lys 645 650 655 Lys Lys Ile Lys
Glu Lys Tyr Ile Asp Gln Glu Glu Leu Asn Lys Thr 660 665 670 Lys Pro
Ile Trp Thr Arg Asn Pro Asp Asp Ile Thr Asn Glu Glu Tyr 675 680 685
Gly Glu Phe Tyr Lys Ser Leu Thr Asn Asp Trp Glu Asp His Leu Ala 690
695 700 Val Lys His Phe Ser Val Glu Gly Gln Leu Glu Phe Arg Ala Leu
Leu 705 710 715 720 Phe Val Pro Arg Arg Ala Pro Phe Asp Leu Phe Glu
Asn Arg Lys Lys 725 730 735 Lys Asn Asn Ile Lys Leu Tyr Val Arg Arg
Val Phe Ile Met Asp Asn 740 745 750 Cys Glu Glu Leu Ile Pro Glu Tyr
Leu Asn Phe Ile Arg Gly Val Val 755 760 765 Asp Ser Glu Asp Leu Pro
Leu Asn Ile Ser Arg Glu Met Leu Gln Gln 770 775 780 Ser Lys Ile Leu
Lys Val Ile Arg Lys Asn Leu Val Lys Lys Cys Leu 785 790 795 800 Glu
Leu Phe Thr Glu Leu Ala Glu Asp Lys Glu Asn Tyr Lys Lys Phe 805 810
815 Tyr Glu Gln Phe Ser Lys Asn Ile Lys Leu Gly Ile His Glu Asp Ser
820 825 830 Gln Asn Arg Lys Lys Leu Ser Glu Leu Leu Arg Tyr Tyr Thr
Ser Ala 835 840 845 Ser Gly Asp Glu Met Val Ser Leu Lys Asp Tyr Cys
Thr Arg Met Lys 850 855 860 Glu Asn Gln Lys His Ile Tyr Tyr Ile Thr
Gly Glu Thr Lys Asp Gln 865 870 875 880 Val Ala Asn Ser Ala Phe Val
Glu Arg Leu Arg Lys His Gly Leu Glu 885 890 895 Val Ile Tyr Met Ile
Glu Pro Ile Asp Glu Tyr Cys Val Gln Gln Leu
900 905 910 Lys Glu Phe Glu Gly Lys Thr Leu Val Ser Val Thr Lys Glu
Gly Leu 915 920 925 Glu Leu Pro Glu Asp Glu Glu Glu Lys Lys Lys Gln
Glu Glu Lys Lys 930 935 940 Thr Lys Phe Glu Asn Leu Cys Lys Ile Met
Lys Asp Ile Leu Glu Lys 945 950 955 960 Lys Val Glu Lys Val Val Val
Ser Asn Arg Leu Val Thr Ser Pro Cys 965 970 975 Cys Ile Val Thr Ser
Thr Tyr Gly Trp Thr Ala Asn Met Glu Arg Ile 980 985 990 Met Lys Ala
Gln Ala Leu Arg Asp Asn Ser Thr Met Gly Tyr Met Ala 995 1000 1005
Ala Lys Lys His Leu Glu Ile Asn Pro Asp His Ser Ile Ile Glu 1010
1015 1020 Thr Leu Arg Gln Lys Ala Glu Ala Asp Lys Asn Asp Lys Ser
Val 1025 1030 1035 Lys Asp Leu Val Ile Leu Leu Tyr Glu Thr Ala Leu
Leu Ser Ser 1040 1045 1050 Gly Phe Ser Leu Glu Asp Pro Gln Thr His
Ala Asn Arg Ile Tyr 1055 1060 1065 Arg Met Ile Lys Leu Gly Leu Gly
Ile Asp Glu Asp Asp Pro Thr 1070 1075 1080 Ala Asp Asp Thr Ser Ala
Ala Val Thr Glu Glu Met Pro Pro Leu 1085 1090 1095 Glu Gly Asp Asp
Asp Thr Ser Arg Met Glu Glu Val Asp 1100 1105 1110
361111PRTArtificial Sequence3E10 Fv-Hsp90 amino acid sequence for a
chimeric protein comprising a single chain Fv antibody fragment
derived from Mus musculus 3E10 hybridoma and Hsp90 from Homo
sapiens joined by human CH1 linker and swivel sequence 36Met Arg
Phe Pro Ser Leu Phe Thr Ala Val Leu Phe Ala Ala Ser Ser 1 5 10 15
Ala Leu Ala Ala Pro Val Asn Thr Thr Thr Glu Asp Glu Thr Ala Gln 20
25 30 Ile Pro Ala Glu Ala Val Ile Gly Tyr Ser Asp Leu Glu Gly Asp
Phe 35 40 45 Asp Val Ala Val Leu Pro Phe Ser Asn Ser Thr Asn Asn
Gly Leu Leu 50 55 60 Phe Ile Asn Thr Thr Ile Ala Ser Ile Ala Ala
Lys Glu Glu Gly Val 65 70 75 80 Ser Leu Glu Lys Arg Glu Ala Glu Ala
Glu Phe His His His His His 85 90 95 His Ala Gly Ile His Asp Ile
Val Leu Thr Gln Ser Pro Ala Ser Leu 100 105 110 Ala Val Ser Leu Gly
Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Lys 115 120 125 Ser Val Ser
Thr Ser Ser Tyr Ser Tyr Met His Trp Tyr Gln Gln Lys 130 135 140 Pro
Gly Gln Pro Pro Lys Leu Leu Ile Lys Tyr Ala Ser Tyr Leu Glu 145 150
155 160 Ser Gly Val Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp
Phe 165 170 175 Thr Leu Asn Ile His Pro Val Glu Glu Glu Asp Ala Ala
Thr Tyr Tyr 180 185 190 Cys Gln His Ser Arg Glu Phe Pro Trp Thr Phe
Gly Gly Gly Thr Lys 195 200 205 Leu Glu Ile Lys Arg Ala Asp Ala Ala
Pro Gly Gly Gly Gly Ser Gly 210 215 220 Gly Gly Gly Ser Gly Gly Gly
Gly Ser Glu Val Gln Leu Val Glu Ser 225 230 235 240 Gly Gly Gly Leu
Val Lys Pro Gly Gly Ser Arg Lys Leu Ser Cys Ala 245 250 255 Ala Ser
Gly Phe Thr Phe Ser Asn Tyr Gly Met His Trp Val Arg Gln 260 265 270
Ala Pro Glu Lys Gly Leu Glu Trp Val Ala Tyr Ile Ser Ser Gly Ser 275
280 285 Ser Thr Ile Tyr Tyr Ala Asp Thr Val Lys Gly Arg Phe Thr Ile
Ser 290 295 300 Arg Asp Asn Ala Lys Asn Thr Leu Phe Leu Gln Met Thr
Ser Leu Arg 305 310 315 320 Ser Glu Asp Thr Ala Met Arg Gly Leu Leu
Leu Asp Tyr Trp Gly Gln 325 330 335 Gly Thr Thr Leu Thr Val Ser Ser
Leu Glu Gln Lys Leu Ile Ser Glu 340 345 350 Glu Asp Leu Asn Ser Ala
Val Asp Ala Ser Thr Lys Gly Pro Ser Val 355 360 365 Phe Pro Leu Ala
Pro Leu Glu Ser Ser Gly Ser Met Pro Glu Glu Thr 370 375 380 Gln Thr
Gln Asp Gln Pro Met Glu Glu Glu Glu Val Glu Thr Phe Ala 385 390 395
400 Phe Gln Ala Glu Ile Ala Gln Leu Met Ser Leu Ile Ile Asn Thr Phe
405 410 415 Tyr Ser Asn Lys Glu Ile Phe Leu Arg Glu Leu Ile Ser Asn
Ser Ser 420 425 430 Asp Ala Leu Asp Lys Ile Arg Tyr Glu Ser Leu Thr
Asp Pro Ser Lys 435 440 445 Leu Asp Ser Gly Lys Glu Leu His Ile Asn
Leu Ile Pro Asn Lys Gln 450 455 460 Asp Arg Thr Leu Thr Ile Val Asp
Thr Gly Ile Gly Met Thr Lys Ala 465 470 475 480 Asp Leu Ile Asn Asn
Leu Gly Thr Ile Ala Lys Ser Gly Thr Lys Ala 485 490 495 Phe Met Glu
Ala Leu Gln Ala Gly Ala Asp Ile Ser Met Ile Gly Gln 500 505 510 Phe
Gly Val Gly Phe Tyr Ser Ala Tyr Leu Val Ala Glu Lys Val Thr 515 520
525 Val Ile Thr Lys His Asn Asp Asp Glu Gln Tyr Ala Trp Glu Ser Ser
530 535 540 Ala Gly Gly Ser Phe Thr Val Arg Thr Asp Thr Gly Glu Pro
Met Gly 545 550 555 560 Arg Gly Thr Lys Val Ile Leu His Leu Lys Glu
Asp Gln Thr Glu Tyr 565 570 575 Leu Glu Glu Arg Arg Ile Lys Glu Ile
Val Lys Lys His Ser Gln Phe 580 585 590 Ile Gly Tyr Pro Ile Thr Leu
Phe Val Glu Lys Glu Arg Asp Lys Glu 595 600 605 Val Ser Asp Asp Glu
Ala Glu Glu Lys Glu Asp Lys Glu Glu Glu Lys 610 615 620 Glu Lys Glu
Glu Lys Glu Ser Glu Asp Lys Pro Glu Ile Glu Asp Val 625 630 635 640
Gly Ser Asp Glu Glu Glu Glu Lys Lys Asp Gly Asp Lys Lys Lys Lys 645
650 655 Lys Lys Ile Lys Glu Lys Tyr Ile Asp Gln Glu Glu Leu Asn Lys
Thr 660 665 670 Lys Pro Ile Trp Thr Arg Asn Pro Asp Asp Ile Thr Asn
Glu Glu Tyr 675 680 685 Gly Glu Phe Tyr Lys Ser Leu Thr Asn Asp Trp
Glu Asp His Leu Ala 690 695 700 Val Lys His Phe Ser Val Glu Gly Gln
Leu Glu Phe Arg Ala Leu Leu 705 710 715 720 Phe Val Pro Arg Arg Ala
Pro Phe Asp Leu Phe Glu Asn Arg Lys Lys 725 730 735 Lys Asn Asn Ile
Lys Leu Tyr Val Arg Arg Val Phe Ile Met Asp Asn 740 745 750 Cys Glu
Glu Leu Ile Pro Glu Tyr Leu Asn Phe Ile Arg Gly Val Val 755 760 765
Asp Ser Glu Asp Leu Pro Leu Asn Ile Ser Arg Glu Met Leu Gln Gln 770
775 780 Ser Lys Ile Leu Lys Val Ile Arg Lys Asn Leu Val Lys Lys Cys
Leu 785 790 795 800 Glu Leu Phe Thr Glu Leu Ala Glu Asp Lys Glu Asn
Tyr Lys Lys Phe 805 810 815 Tyr Glu Gln Phe Ser Lys Asn Ile Lys Leu
Gly Ile His Glu Asp Ser 820 825 830 Gln Asn Arg Lys Lys Leu Ser Glu
Leu Leu Arg Tyr Tyr Thr Ser Ala 835 840 845 Ser Gly Asp Glu Met Val
Ser Leu Lys Asp Tyr Cys Thr Arg Met Lys 850 855 860 Glu Asn Gln Lys
His Ile Tyr Tyr Ile Thr Gly Glu Thr Lys Asp Gln 865 870 875 880 Val
Ala Asn Ser Ala Phe Val Glu Arg Leu Arg Lys His Gly Leu Glu 885 890
895 Val Ile Tyr Met Ile Glu Pro Ile Asp Glu Tyr Cys Val Gln Gln Leu
900 905 910 Lys Glu Phe Glu Gly Lys Thr Leu Val Ser Val Thr Lys Glu
Gly Leu 915 920 925 Glu Leu Pro Glu Asp Glu Glu Glu Lys Lys Lys Gln
Glu Glu Lys Lys 930 935 940 Thr Lys Phe Glu Asn Leu Cys Lys Ile Met
Lys Asp Ile Leu Glu Lys 945 950 955 960 Lys Val Glu Lys Val Val Val
Ser Asn Arg Leu Val Thr Ser Pro Cys 965 970 975 Cys Ile Val Thr Ser
Thr Tyr Gly Trp Thr Ala Asn Met Glu Arg Ile 980 985 990 Met Lys Ala
Gln Ala Leu Arg Asp Asn Ser Thr Met Gly Tyr Met Ala 995 1000 1005
Ala Lys Lys His Leu Glu Ile Asn Pro Asp His Ser Ile Ile Glu 1010
1015 1020 Thr Leu Arg Gln Lys Ala Glu Ala Asp Lys Asn Asp Lys Ser
Val 1025 1030 1035 Lys Asp Leu Val Ile Leu Leu Tyr Glu Thr Ala Leu
Leu Ser Ser 1040 1045 1050 Gly Phe Ser Leu Glu Asp Pro Gln Thr His
Ala Asn Arg Ile Tyr 1055 1060 1065 Arg Met Ile Lys Leu Gly Leu Gly
Ile Asp Glu Asp Asp Pro Thr 1070 1075 1080 Ala Asp Asp Thr Ser Ala
Ala Val Thr Glu Glu Met Pro Pro Leu 1085 1090 1095 Glu Gly Asp Asp
Asp Thr Ser Arg Met Glu Glu Val Asp 1100 1105 1110
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