实施例7:抗PCSK9单抗抑制PCSK9诱导HEK293细胞摄取LDL-C能力的下调。
本实施例研究用悬浮培养于293Freestyle培养基(Invitrogen)的HEK293细胞。将HEK293细胞密度调整至8*10E5cells/ml,接种于24孔板(500ul/孔)。然后按照表5所列方式处理细胞,悬浮培养2.5h。然后再分别在每个孔中补加BODIPYFLLDL(Invitrogen,Cat#:L3483)至终浓度6ug/ml,继续悬浮培养2.5h。然后分别收集各孔中细胞,用PBS洗涤2次后,用BD公司的AccuriC6流式细胞仪分析BODIPY信号(激发波长488nM,发射波长520)。以未处理的HEK293细胞为对照,比较各种处理后细胞摄取LDL-C的水平。结果(图5)显示:10ug/ml的PCSK9处理HEK293细胞后导致细胞摄取LDL-C量明显下降,而多种抗PCSK9单抗(20ug/ml)能够明显抑制PCSK9诱导的HEK293细胞摄取LDL-C水平的下调。
表5:HEK293细胞的不同处理方式。
序列表
<110>北京智仁美博生物科技有限公司
<120>抗人PCSK9单克隆抗体
<160>24
<210>1
<211>639
<212>PRT
<213>智人(Homosapiens)
<220>
<223>huPCSK9
<400>1
GluAlaProGluHisGlyThrThrAlaThrPheHisArgCysAlaLys
151015
AspProTrpArgLeuProGlyThrTyrValValValLeuLysGluGlu
202530
ThrHisLeuSerGlnSerGluArgThrAlaArgArgLeuGlnAlaGln
354045
AlaAlaArgArgGlyTyrLeuThrLysIleLeuHisValPheHisGly
505560
LeuLeuProGlyPheLeuValLysMetSerGlyAspLeuLeuGluLeu
65707580
AlaLeuLysLeuProHisValAspTyrIleGluGluAspSerSerVal
859095
PheAlaGlnSerIleProTrpAsnLeuGluArgIleThrProProArg
100105110
TyrArgAlaAspGluTyrGlnProProAspGlyGlySerLeuValGlu
115120125
ValTyrLeuLeuAspThrSerIleGlnSerAspHisArgGluIleGlu
130135140
GlyArgValMetValThrAspPheGluAsnValProGluGluAspGly
145150155160
ThrArgPheHisArgGlnAlaSerLysCysAspSerHisGlyThrHis
165170175
LeuAlaGlyValValSerGlyArgAspAlaGlyValAlaLysGlyAla
180185190
SerMetArgSerLeuArgValLeuAsnCysGlnGlyLysGlyThrVal
195200205
SerGlyThrLeuIleGlyLeuGluPheIleArgLysSerGlnLeuVal
210215220
GlnProValGlyProLeuValValLeuLeuProLeuAlaGlyGlyTyr
225230235240
SerArgValLeuAsnAlaAlaCysGlnArgLeuAlaArgAlaGlyVal
245250255
ValLeuValThrAlaAlaGlyAsnPheArgAspAspAlaCysLeuTyr
260265270
SerProAlaSerAlaProGluValIleThrValGlyAlaThrAsnAla
275280285
GlnAspGlnProValThrLeuGlyThrLeuGlyThrAsnPheGlyArg
290295300
CysValAspLeuPheAlaProGlyGluAspIleIleGlyAlaSerSer
305310315320
AspCysSerThrCysPheValSerGlnSerGlyThrSerGlnAlaAla
325330335
AlaHisValAlaGlyIleAlaAlaMetMetLeuSerAlaGluProGlu
340345350
LeuThrLeuAlaGluLeuArgGlnArgLeuIleHisPheSerAlaLys
355360365
AspValIleAsnGluAlaTrpPheProGluAspGlnArgValLeuThr
370375380
ProAsnLeuValAlaAlaLeuProProSerThrHisGlyAlaGlyTrp
385390395400
GlnLeuPheCysArgThrValTrpSerAlaHisSerGlyProThrArg
405410415
MetAlaThrAlaIleAlaArgCysAlaProAspGluGluLeuLeuSer
420425430
CysSerSerPheSerArgSerGlyLysArgArgGlyGluArgMetGlu
435440445
AlaGlnGlyGlyLysLeuValCysArgAlaHisAsnAlaPheGlyGly
450455460
GluGlyValTyrAlaIleAlaArgCysCysLeuLeuProGlnAlaAsn
465470475480
CysSerValHisThrAlaProProAlaGluAlaSerMetGlyThrArg
485490495
ValHisCysHisGlnGlnGlyHisValLeuThrGlyCysSerSerHis
500505510
TrpGluValGluAspLeuGlyThrHisLysProProValLeuArgPro
515520525
ArgGlyGlnProAsnGlnCysValGlyHisArgGluAlaSerIleHis
530535540
AlaSerCysCysHisAlaProGlyLeuGluCysLysValLysGluHis
545550555560
GlyIleProAlaProGlnGluGlnValThrValAlaCysGluGluGly
565570575
TrpThrLeuThrGlyCysSerAlaLeuProGlyThrSerHisValLeu
580585590
GlyAlaTyrAlaValAspAsnThrCysValValArgSerArgAspVal
595600605
SerThrThrGlySerThrSerGluGluAlaValThrAlaValAlaIle
610615620
CysCysArgSerArgHisLeuAlaGlnAlaSerGlnGluLeuGln
625630635
<210>2
<211>660
<212>PRT
<213>小鼠(Musmusculus)
<220>
<223>mopcsk9
<400>2
GlnAspGluAspGlyAspTyrGluGluLeuMetLeuAlaLeuProSer
151015
GlnGluAspGlyLeuAlaAspGluAlaAlaHisValAlaThrAlaThr
202530
PheArgArgCysSerLysGluAlaTrpArgLeuProGlyThrTyrIle
354045
ValValLeuMetGluGluThrGlnArgLeuGlnIleGluGlnThrAla
505560
HisArgLeuGlnThrArgAlaAlaArgArgGlyTyrValIleLysVal
65707580
LeuHisIlePheTyrAspLeuPheProGlyPheLeuValLysMetSer
859095
SerAspLeuLeuGlyLeuAlaLeuLysLeuProHisValGluTyrIle
100105110
GluGluAspSerPheValPheAlaGlnSerIleProTrpAsnLeuGlu
115120125
ArgIleIleProAlaTrpHisGlnThrGluGluAspArgSerProAsp
130135140
GlySerSerGlnValGluValTyrLeuLeuAspThrSerIleGlnGly
145150155160
AlaHisArgGluIleGluGlyArgValThrIleThrAspPheAsnSer
165170175
ValProGluGluAspGlyThrArgPheHisArgGlnAlaSerLysCys
180185190
AspSerHisGlyThrHisLeuAlaGlyValValSerGlyArgAspAla
195200205
GlyValAlaLysGlyThrSerLeuHisSerLeuArgValLeuAsnCys
210215220
GlnGlyLysGlyThrValSerGlyThrLeuIleGlyLeuGluPheIle
225230235240
ArgLysSerGlnLeuIleGlnProSerGlyProLeuValValLeuLeu
245250255
ProLeuAlaGlyGlyTyrSerArgIleLeuAsnAlaAlaCysArgHis
260265270
LeuAlaArgThrGlyValValLeuValAlaAlaAlaGlyAsnPheArg
275280285
AspAspAlaCysLeuTyrSerProAlaSerAlaProGluValIleThr
290295300
ValGlyAlaThrAsnAlaGlnAspGlnProValThrLeuGlyThrLeu
305310315320
GlyThrAsnPheGlyArgCysValAspLeuPheAlaProGlyLysAsp
325330335
IleIleGlyAlaSerSerAspCysSerThrCysPheMetSerGlnSer
340345350
GlyThrSerGlnAlaAlaAlaHisValAlaGlyIleValAlaArgMet
355360365
LeuSerArgGluProThrLeuThrLeuAlaGluLeuArgGlnArgLeu
370375380
IleHisPheSerThrLysAspValIleAsnMetAlaTrpPheProGlu
385390395400
AspGlnGlnValLeuThrProAsnLeuValAlaThrLeuProProSer
405410415
ThrHisGluThrGlyGlyGlnLeuLeuCysArgThrValTrpSerAla
420425430
HisSerGlyProThrArgThrAlaThrAlaThrAlaArgCysAlaPro
435440445
GluGluGluLeuLeuSerCysSerSerPheSerArgSerGlyArgArg
450455460
ArgGlyAspTrpIleGluAlaIleGlyGlyGlnGlnValCysLysAla
465470475480
LeuAsnAlaPheGlyGlyGluGlyValTyrAlaValAlaArgCysCys
485490495
LeuValProArgAlaAsnCysSerIleHisAsnThrProAlaAlaArg
500505510
AlaGlyLeuGluThrHisValHisCysHisGlnLysAspHisValLeu
515520525
ThrGlyCysSerPheHisTrpGluValGluAspLeuSerValArgArg
530535540
GlnProAlaLeuArgSerArgArgGlnProGlyGlnCysValGlyHis
545550555560
GlnAlaAlaSerValTyrAlaSerCysCysHisAlaProGlyLeuGlu
565570575
CysLysIleLysGluHisGlyIleSerGlyProSerGluGlnValThr
580585590
ValAlaCysGluAlaGlyTrpThrLeuThrGlyCysAsnValLeuPro
595600605
GlyAlaSerLeuThrLeuGlyAlaTyrSerValAspAsnLeuCysVal
610615620
AlaArgValHisAspThrAlaArgAlaAspArgThrSerGlyGluAla
625630635640
ThrValAlaAlaAlaIleCysCysArgSerArgProSerAlaLysAla
645650655
SerTrpValGln
660
<210>3
<211>639
<212>PRT
<213>猕猴(Macacamulatta)
<220>
<223>mmPCSK9
<400>3
AspAlaProGluHisGlyAlaThrAlaThrPheHisArgCysAlaLys
151015
AspProTrpArgLeuProGlyThrTyrValValValLeuLysGluGlu
202530
ThrHisArgSerGlnSerGluArgThrAlaArgArgLeuGlnAlaGln
354045
AlaAlaArgArgGlyTyrLeuThrLysIleLeuHisValPheHisHis
505560
LeuLeuProGlyPheLeuValLysMetSerGlyAspLeuLeuGluLeu
65707580
AlaLeuLysLeuProHisValAspTyrIleGluGluAspSerSerVal
859095
PheAlaGlnSerIleProTrpAsnLeuGluArgIleThrProAlaArg
100105110
TyrArgAlaAspGluTyrGlnProProLysGlyGlySerLeuValGlu
115120125
ValTyrLeuLeuAspThrSerIleGlnSerAspHisArgGluIleGlu
130135140
GlyArgValMetValThrAspPheGluSerValProGluGluAspGly
145150155160
ThrArgPheHisArgGlnAlaSerLysCysAspSerHisGlyThrHis
165170175
LeuAlaGlyValValSerGlyArgAspAlaGlyValAlaLysGlyAla
180185190
GlyLeuArgSerLeuArgValLeuAsnCysGlnGlyLysGlyThrVal
195200205
SerGlyThrLeuIleGlyLeuGluPheIleArgLysSerGlnLeuVal
210215220
GlnProValGlyProLeuValValLeuLeuProLeuAlaGlyGlyTyr
225230235240
SerArgValPheAsnAlaAlaCysGlnArgLeuAlaArgAlaGlyVal
245250255
ValLeuValThrAlaAlaGlyAsnPheArgAspAspAlaCysLeuTyr
260265270
SerProAlaSerAlaProGluValIleThrValGlyAlaThrAsnAla
275280285
GlnAspGlnProValThrLeuGlyThrLeuGlyThrAsnPheGlyArg
290295300
CysValAspLeuPheAlaProGlyGluAspIleIleGlyAlaSerSer
305310315320
AspCysSerThrCysPheValSerArgSerGlyThrSerGlnAlaAla
325330335
AlaHisValAlaGlyIleAlaAlaMetMetLeuSerAlaGluProGlu
340345350
LeuThrLeuAlaGluLeuArgGlnArgLeuIleHisPheSerAlaLys
355360365
AspValIleAsnGluAlaTrpPheProGluAspGlnArgValLeuThr
370375380
ProAsnLeuValAlaAlaLeuProProSerThrHisArgAlaGlyTrp
385390395400
GlnLeuPheCysArgThrValTrpSerAlaHisSerGlyProThrArg
405410415
MetAlaThrAlaValAlaArgCysAlaGlnAspGluGluLeuLeuSer
420425430
CysSerSerPheSerArgSerGlyLysArgArgGlyGluArgIleGlu
435440445
AlaGlnGlyGlyLysArgValCysArgAlaHisAsnAlaPheGlyGly
450455460
GluGlyValTyrAlaIleAlaArgCysCysLeuLeuProGlnValAsn
465470475480
CysSerValHisThrAlaProProAlaGlyAlaSerMetGlyThrArg
485490495
ValHisCysHisGlnGlnGlyHisValLeuThrGlyCysSerSerHis
500505510
TrpGluValGluAspLeuGlyThrHisLysProProValLeuArgPro
515520525
ArgGlyGlnProAsnGlnCysValGlyHisArgGluAlaSerIleHis
530535540
AlaSerCysCysHisAlaProGlyLeuGluCysLysValLysGluHis
545550555560
GlyIleProAlaProGlnGluGlnValIleValAlaCysGluAspGly
565570575
TrpThrLeuThrGlyCysSerProLeuProGlyThrSerHisValLeu
580585590
GlyAlaTyrAlaValAspAsnThrCysValValArgSerArgAspVal
595600605
SerThrThrGlySerThrSerLysGluAlaValAlaAlaValAlaIle
610615620
CysCysArgSerArgHisLeuValGlnAlaSerGlnGluLeuGln
625630635
<210>4
<211>120
<212>PRT
<213>智人(Homosapiens)
<220>
<223>LDLR-EGFAB
<400>4
ThrLeuCysGluGlyProAsnLysPheLysCysHisSerGlyGluCys
151015
IleThrLeuAspLysValCysAsnMetAlaArgAspCysArgAspTrp
202530
SerAspGluProIleLysGluCysGlyThrAsnGluCysLeuAspAsn
354045
AsnGlyGlyCysSerHisValCysAsnAspLeuLysIleGlyTyrGlu
505560
CysLeuCysProAspGlyPheGlnLeuValAlaGlnArgArgCysGlu
65707580
AspIleAspGluCysGlnAspProAspThrCysSerGlnLeuCysVal
859095
AsnLeuGluGlyGlyTyrLysCysGlnCysGluGluGlyPheGlnLeu
100105110
AspProHisThrLysAlaCysLys
115120
<210>5
<211>11
<212>PRT
<213>人工序列
<220>
<223>his标签
<400>5
AlaSerGlyAlaAlaHisHisHisHisHisHis
1510
<210>6
<211>234
<212>PRT
<213>小鼠(Musmusculus)
<220>
<223>mFc标签
<400>6
AlaSerProArgGlyProThrIleLysProCysProProCysLysCys
151015
ProAlaProAsnLeuLeuGlyGlyProSerValPheIlePheProPro
202530
LysIleLysAspValLeuMetIleSerLeuSerProIleValThrCys
354045
ValValValAspValSerGluAspAspProAspValGlnIleSerTrp
505560
PheValAsnAsnValGluValHisThrAlaGlnThrGlnThrHisArg
65707580
GluAspTyrAsnSerThrLeuArgValValSerAlaLeuProIleGln
859095
HisGlnAspTrpMetSerGlyLysGluPheLysCysLysValAsnAsn
100105110
LysAspLeuProAlaProIleGluArgThrIleSerLysProLysGly
115120125
SerValArgAlaProGlnValTyrValLeuProProProGluGluGlu
130135140
MetThrLysLysGlnValThrLeuThrCysMetValThrAspPheMet
145150155160
ProGluAspIleTyrValGluTrpThrAsnAsnGlyLysThrGluLeu
165170175
AsnTyrLysAsnThrGluProValLeuAspSerAspGlySerTyrPhe
180185190
MetTyrSerLysLeuArgValGluLysLysAsnTrpValGluArgAsn
195200205
SerTyrSerCysSerValValHisGluGlyLeuHisAsnHisHisThr
210215220
ThrLysSerPheSerArgThrProGlyLys
225230
<210>7
<211>330
<212>PRT
<213>智人(Homosapiens)
<220>
<223>IgG-C1
<400>7
AlaSerThrLysGlyProSerValPheProLeuAlaProSerSerLys
151015
SerThrSerGlyGlyThrAlaAlaLeuGlyCysLeuValLysAspTyr
202530
PheProGluProValThrValSerTrpAsnSerGlyAlaLeuThrSer
354045
GlyValHisThrPheProAlaValLeuGlnSerSerGlyLeuTyrSer
505560
LeuSerSerValValThrValProSerSerSerLeuGlyThrGlnThr
65707580
TyrIleCysAsnValAsnHisLysProSerAsnThrLysValAspLys
859095
ArgValGluProLysSerCysAspLysThrHisThrCysProProCys
100105110
ProAlaProGluLeuLeuGlyGlyProSerValPheLeuPheProPro
115120125
LysProLysAspThrLeuMetIleSerArgThrProGluValThrCys
130135140
ValValValAspValSerHisGluAspProGluValLysPheAsnTrp
145150155160
TyrValAspGlyValGluValHisAsnAlaLysThrLysProArgGlu
165170175
GluGlnTyrAsnSerThrTyrArgValValSerValLeuThrValLeu
180185190
HisGlnAspTrpLeuAsnGlyLysGluTyrLysCysLysValSerAsn
195200205
LysAlaLeuProAlaProIleGluLysThrIleSerLysAlaLysGly
210215220
GlnProArgGluProGlnValTyrThrLeuProProSerArgGluGlu
225230235240
MetThrLysAsnGlnValSerLeuThrCysLeuValLysGlyPheTyr
245250255
ProSerAspIleAlaValGluTrpGluSerAsnGlyGlnProGluAsn
260265270
AsnTyrLysThrThrProProValLeuAspSerAspGlySerPhePhe
275280285
LeuTyrSerLysLeuThrValAspLysSerArgTrpGlnGlnGlyAsn
290295300
ValPheSerCysSerValMetHisGluAlaLeuHisAsnHisTyrThr
305310315320
GlnLysSerLeuSerLeuSerProGlyLys
325330
<210>8
<211>326
<212>PRT
<213>智人(Homosapiens)
<220>
<223>IgG2-C
<400>8
AlaSerThrLysGlyProSerValPheProLeuAlaProCysSerArg
151015
SerThrSerGluSerThrAlaAlaLeuGlyCysLeuValLysAspTyr
202530
PheProGluProValThrValSerTrpAsnSerGlyAlaLeuThrSer
354045
GlyValHisThrPheProAlaValLeuGlnSerSerGlyLeuTyrSer
505560
LeuSerSerValValThrValProSerSerAsnPheGlyThrGlnThr
65707580
TyrThrCysAsnValAspHisLysProSerAsnThrLysValAspLys
859095
ThrValGluArgLysCysCysValGluCysProProCysProAlaPro
100105110
ProValAlaGlyProSerValPheLeuPheProProLysProLysAsp
115120125
ThrLeuMetIleSerArgThrProGluValThrCysValValValAsp
130135140
ValSerHisGluAspProGluValGlnPheAsnTrpTyrValAspGly
145150155160
ValGluValHisAsnAlaLysThrLysProArgGluGluGlnPheAsn
165170175
SerThrPheArgValValSerValLeuThrValValHisGlnAspTrp
180185190
LeuAsnGlyLysGluTyrLysCysLysValSerAsnLysGlyLeuPro
195200205
AlaProIleGluLysThrIleSerLysThrLysGlyGlnProArgGlu
210215220
ProGlnValTyrThrLeuProProSerArgGluGluMetThrLysAsn
225230235240
GlnValSerLeuThrCysLeuValLysGlyPheTyrProSerAspIle
245250255
AlaValGluTrpGluSerAsnGlyGlnProGluAsnAsnTyrLysThr
260265270
ThrProProMetLeuAspSerAspGlySerPhePheLeuTyrSerLys
275280285
LeuThrValAspLysSerArgTrpGlnGlnGlyAsnValPheSerCys
290295300
SerValMetHisGluAlaLeuHisAsnHisTyrThrGlnLysSerLeu
305310315320
SerLeuSerProGlyLys
325
<210>9
<211>327
<212>PRT
<213>智人(Homosapiens)
<220>
<223>IgG4-C
<400>9
AlaSerThrLysGlyProSerValPheProLeuAlaProCysSerArg
151015
SerThrSerGluSerThrAlaAlaLeuGlyCysLeuValLysAspTyr
202530
PheProGluProValThrValSerTrpAsnSerGlyAlaLeuThrSer
354045
GlyValHisThrPheProAlaValLeuGlnSerSerGlyLeuTyrSer
505560
LeuSerSerValValThrValProSerSerSerLeuGlyThrLysThr
65707580
TyrThrCysAsnValAspHisLysProSerAsnThrLysValAspLys
859095
ArgValGluSerLysTyrGlyProProCysProProCysProAlaPro
100105110
GluPheLeuGlyGlyProSerValPheLeuPheProProLysProLys
115120125
AspThrLeuMetIleSerArgThrProGluValThrCysValValVal
130135140
AspValSerGlnGluAspProGluValGlnPheAsnTrpTyrValAsp
145150155160
GlyValGluValHisAsnAlaLysThrLysProArgGluGluGlnPhe
165170175
AsnSerThrTyrArgValValSerValLeuThrValLeuHisGlnAsp
180185190
TrpLeuAsnGlyLysGluTyrLysCysLysValSerAsnLysGlyLeu
195200205
ProSerSerIleGluLysThrIleSerLysAlaLysGlyGlnProArg
210215220
GluProGlnValTyrThrLeuProProSerGlnGluGluMetThrLys
225230235240
AsnGlnValSerLeuThrCysLeuValLysGlyPheTyrProSerAsp
245250255
IleAlaValGluTrpGluSerAsnGlyGlnProGluAsnAsnTyrLys
260265270
ThrThrProProValLeuAspSerAspGlySerPhePheLeuTyrSer
275280285
ArgLeuThrValAspLysSerArgTrpGlnGluGlyAsnValPheSer
290295300
CysSerValMetHisGluAlaLeuHisAsnHisTyrThrGlnLysSer
305310315320
LeuSerLeuSerLeuGlyLys
325
<210>10
<211>107
<212>PRT
<213>智人(Homosapiens)
<220>
<223>kappa亚型轻链恒定区
<400>10
ArgThrValAlaAlaProSerValPheIlePheProProSerAspGlu
151015
GlnLeuLysSerGlyThrAlaSerValValCysLeuLeuAsnAsnPhe
202530
TyrProArgGluAlaLysValGlnTrpLysValAspAsnAlaLeuGln
354045
SerGlyAsnSerGlnGluSerValThrGluGlnAspSerLysAspSer
505560
ThrTyrSerLeuSerSerThrLeuThrLeuSerLysAlaAspTyrGlu
65707580
LysHisLysValTyrAlaCysGluValThrHisGlnGlyLeuSerSer
859095
ProValThrLysSerPheAsnArgGlyGluCys
100105
<210>11
<211>105
<212>PRT
<213>智人(Homosapiens)
<220>
<223>lambda亚型轻链恒定区
<400>11
GlnProLysAlaAlaProSerValThrLeuPheProProSerSerGlu
151015
GluLeuGlnAlaAsnLysAlaThrLeuValCysLeuIleSerAspPhe
202530
TyrProGlyAlaValThrValAlaTrpLysAlaAspSerSerProVal
354045
LysAlaGlyValGluThrThrThrProSerLysGlnSerAsnAsnLys
505560
TyrAlaAlaSerSerTyrLeuSerLeuThrProGluGlnTrpLysSer
65707580
HisArgSerTyrSerCysGlnValThrHisGluGlySerThrValGlu
859095
LysThrValAlaProThrGluCysSer
100105
<210>12
<211>245
<212>PRT
<213>人工序列
<220>
<223>S2B8
<400>12
GlnValGlnLeuValGluSerGlyGlyGlyLeuValLysProGlyGly
151015
SerLeuArgLeuSerCysAlaValSerGlyPheThrPheSerArgHis
202530
SerMetAsnTrpValArgGlnAlaProGlyLysGlyLeuGluTrpVal
354045
SerSerIleSerSerGlySerSerAsnIleTyrTyrAlaAspSerVal
505560
LysGlyArgPheThrIleSerArgAspAsnAlaLysAsnSerLeuTyr
65707580
LeuGlnMetAsnSerLeuArgAspGluAspThrAlaValTyrTyrCys
859095
AlaArgGlyGlyLeuGlyProGluTyrLeuGlnHisTrpGlyGlnGly
100105110
ThrLeuValThrValSerSerGlyGlyGlyGlySerGlyGlyGlyGly
115120125
SerGlyGlyGlyGlySerSerSerGluLeuThrGlnProAlaSerVal
130135140
SerGlySerProGlyGlnSerIleThrIleSerCysThrGlyThrSer
145150155160
SerAspIleGlyAspAsnAsnPheValSerTrpTyrGlnGlnTyrPro
165170175
GlyLysAlaProLysLeuMetIleTyrAspValSerAsnArgProSer
180185190
GlyValSerAsnArgPheSerGlySerLysSerGlyAsnThrAlaSer
195200205
LeuThrIleSerGlyLeuGlnAlaGluAspGluAlaAspTyrTyrCys
210215220
SerSerTyrThrSerSerSerThrLeuValPheGlyGlyGlyThrLys
225230235240
ValThrValLeuGly
245
<210>13
<211>242
<212>PRT
<213>人工序列
<220>
<223>S2F5
<400>13
GlnValGlnLeuValGlnSerGlyAlaGluValLysLysProGlySer
151015
SerValLysValSerCysLysAlaSerGlyGlyThrPheThrGlyTyr
202530
SerIleHisTrpValArgGlnAlaProGlyGlnGlyLeuGluTrpMet
354045
GlyTrpIleAsnProIleSerGlySerAlaAsnTyrAlaGlnLysPhe
505560
GlnGlyArgValThrIleThrAlaAspLysSerThrSerThrAlaTyr
65707580
MetGluLeuSerSerLeuArgSerGluAspThrAlaValTyrTyrCys
859095
AlaArgThrGlnAsnAsnThrGlyTyrArgGlyThrAspTyrTrpGly
100105110
GlnGlyThrLeuValThrValSerSerGlyGlyGlyGlySerGlyGly
115120125
GlyGlySerGlyGlyGlySerAspIleGlnMetThrGlnSerProSer
130135140
SerLeuSerAlaSerValGlyAspArgValThrIleThrCysArgAla
145150155160
SerGlnSerValSerSerTyrLeuAsnTrpTyrGlnGlnLysProGly
165170175
LysAlaProLysLeuLeuIleTyrGlyAlaSerThrArgProSerGly
180185190
ValProSerArgPheSerGlySerGlySerGlyThrAspPheThrLeu
195200205
ThrIleSerSerLeuGlnProGluAspPheAlaThrTyrTyrCysGln
210215220
GlnHisAlaAspThrProThrThrPheGlyGlnGlyThrLysValGlu
225230235240
IleLys
<210>14
<211>237
<212>PRT
<213>人工序列
<220>
<223>S2G1
<400>14
GlnMetGlnLeuValGlnSerGlyAlaGluValLysLysProGlySer
151015
SerValLysValSerCysLysThrSerGlyGlyThrPheGlySerSer
202530
AlaIleHisTrpValArgGlnAlaProArgGlnGlyLeuGluTrpMet
354045
GlyArgIleAsnProValLeuGlyThrThrAsnTyrThrGlnLysPhe
505560
HisAspArgLeuThrIleThrAlaAspGluSerThrThrThrAlaTyr
65707580
MetGluLeuArgSerLeuArgSerAspAspThrAlaValTyrTyrCys
859095
AlaArgGluGlyIleProMetIleTrpGlyGlnGlyThrMetValThr
100105110
ValSerSerGlyGlyGlyGlySerGlyGlyGlyGlySerGlyGlyGly
115120125
GlySerAspIleGlnMetThrGlnSerProSerSerLeuSerAlaSer
130135140
ValGlyAspArgValThrIleThrCysArgAlaSerGlnGlyIleHis
145150155160
AsnTrpLeuAsnTrpTyrGlnGlnLysProGlyLysAlaProLysLeu
165170175
LeuIleTyrAlaAlaSerAsnArgAlaSerGlyValProSerArgPhe
180185190
SerGlySerGlySerGlyThrAspPheThrLeuThrIleSerSerLeu
195200205
GlnProGluAspPheAlaThrTyrTyrCysGlnGlnSerGlyLysGln
210215220
ProValThrPheGlyGlnGlyThrLysValGluIleLys
225230235
<210>15
<211>235
<212>PRT
<213>人工序列
<220>
<223>S3A12
<400>15
GlnValGlnLeuValGlnSerGlyAlaGluValLysLysProGlyAla
151015
SerValLysValSerCysLysAlaSerGlyTyrAlaIleTyrAsnTyr
202530
GlyIleSerTrpValArgGlnAlaProGlyGlnGlyLeuGluTrpMet
354045
GlyTrpValSerTyrPheGlyGlySerThrThrTyrAlaGlnLysLeu
505560
GlnGlyArgGlyThrMetThrThrAspProSerThrSerThrAlaTyr
65707580
MetGluLeuArgSerLeuArgSerAspAspThrAlaValTyrTyrCys
859095
AlaLysGlyTyrGlyMetGluIleTrpGlyGlnGlyThrThrValSer
100105110
SerGlyGlyGlyGlySerGlyGlyGlyGlySerGlyGlyGlyGlySer
115120125
AspIleGlnMetThrGlnSerProSerSerLeuSerAlaSerValGly
130135140
AspArgValThrIleThrCysArgAlaSerGlnAsnValAsnAsnTrp
145150155160
LeuAspTrpTyrGlnGlnLysProGlyLysAlaProLysLeuLeuIle
165170175
TyrAlaAlaSerThrArgAlaSerGlyValProSerArgPheSerGly
180185190
SerGlySerGlyThrAspPheThrLeuThrIleSerSerLeuGlnPro
195200205
GluAspPheAlaThrTyrTyrCysGlnGlnAspArgAspValProVal
210215220
ThrPheGlyGlnGlyThrLysValGluIleLys
225230235
<210>16
<211>115
<212>PRT
<213>人工序列
<220>
<223>H4H4
<400>16
GlnValGlnLeuValGlnSerGlyAlaGluValLysLysProGlyAla
151015
SerValLysValSerCysLysAlaSerGlyTyrAlaIleTyrAsnTyr
202530
GlyIleSerTrpValArgGlnAlaProGlyGlnGlyLeuGluTrpMet
354045
GlyTrpValSerPheTyrAsnGlyAsnThrAsnTyrAlaGlnLysLeu
505560
GlnGlyArgGlyThrMetThrThrAspProSerThrSerThrAlaTyr
65707580
MetGluLeuArgSerLeuArgSerAspAspThrAlaValTyrTyrCys
859095
AlaArgGlyTyrValMetAspIleTrpGlyGlnGlyThrThrValThr
100105110
ValSerSer
115
<210>17
<211>115
<212>PRT
<213>人工序列
<220>
<223>H7B12
<400>17
GlnValGlnLeuValGlnSerGlyAlaGluValLysLysProGlyAla
151015
SerValLysValSerCysLysAlaSerGlyTyrThrLeuAsnAsnTyr
202530
GlyIleSerTrpValArgGlnAlaProGlyGlnGlyLeuGluTrpMet
354045
GlyTrpValSerPheTyrAsnGlyAsnThrAsnTyrAlaGlnLysLeu
505560
GlnGlyArgGlyThrMetThrThrAspProSerThrSerThrAlaTyr
65707580
MetGluLeuArgSerLeuArgSerAspAspThrAlaValTyrTyrCys
859095
AlaArgGlyTyrValMetAspIleTrpGlyGlnGlyThrThrValThr
100105110
ValSerSer
115
<210>18
<211>115
<212>PRT
<213>人工序列
<220>
<223>H8H7
<400>18
GlnValGlnLeuValGlnSerGlyAlaGluValLysLysProGlyAla
151015
SerValLysValSerCysLysAlaSerGlyTyrAlaIleTyrAsnTyr
202530
GlyIleSerTrpValArgGlnAlaProGlyGlnGlyLeuGluTrpMet
354045
GlyTrpValSerPheTyrAsnGlyAsnThrAsnTyrAlaGlnLysLeu
505560
GlnGlyArgGlyThrMetThrThrAspProSerThrSerThrAlaTyr
65707580
MetGluLeuArgSerLeuArgSerAspAspThrAlaValTyrTyrCys
859095
AlaArgGlyTyrValMetAspPheTrpGlyGlnGlyThrThrValThr
100105110
ValSerSer
115
<210>19
<211>115
<212>PRT
<213>人工序列
<220>
<223>H8F4
<400>19
GlnValGlnLeuValGlnSerGlyAlaGluValLysLysProGlyAla
151015
SerValLysValSerCysLysAlaSerGlyTyrAlaLeuTyrAsnTyr
202530
GlyIleSerTrpValArgGlnAlaProGlyGlnGlyLeuGluTrpMet
354045
GlyTrpValSerPheTyrAsnGlyAsnThrAsnTyrAlaGlnLysLeu
505560
GlnGlyArgGlyThrMetThrThrAspProSerThrSerThrAlaTyr
65707580
MetGluLeuArgSerLeuArgSerAspAspThrAlaValTyrTyrCys
859095
AlaArgGlyTyrValMetAspIleTrpGlyGlnGlyThrThrValThr
100105110
ValSerSer
115
<210>20
<211>108
<212>PRT
<213>人工序列
<220>
<223>L3A8
<400>20
AspIleGlnMetThrGlnSerProSerSerLeuSerAlaSerValGly
151015
AspArgValThrIleThrCysArgAlaSerGlnSerIleAsnAsnTrp
202530
LeuAspTrpTyrGlnGlnLysProGlyLysAlaProLysLeuLeuIle
354045
TyrAlaAlaSerThrArgProSerGlyValProSerArgPheSerGly
505560
SerGlySerGlyThrAspPheThrLeuThrIleSerSerLeuGlnPro
65707580
GluAspPheAlaThrTyrTyrCysGlnGlnAspGlnAspIleProPro
859095
ThrPheGlyGlnGlyThrLysValGluIleLysArg
100105
<210>21
<211>115
<212>PRT
<213>人工序列
<220>
<223>EvoloVH
<400>21
GlnValGlnLeuValGlnSerGlyAlaGluValLysLysProGlyAla
151015
SerValLysValSerCysLysAlaSerGlyTyrThrLeuThrSerTyr
202530
GlyIleSerTrpValArgGlnAlaProGlyGlnGlyLeuGluTrpMet
354045
GlyTrpValSerPheTyrAsnGlyAsnThrAsnTyrAlaGlnLysLeu
505560
GlnGlyArgGlyThrMetThrThrAspProSerThrSerThrAlaTyr
65707580
MetGluLeuArgSerLeuArgSerAspAspThrAlaValTyrTyrCys
859095
AlaArgGlyTyrGlyMetAspValTrpGlyGlnGlyThrThrValThr
100105110
ValSerSer
115
<210>22
<211>109
<212>PRT
<213>人工序列
<220>
<223>EvoloVL
<400>22
GlnSerAlaLeuThrGlnProAlaSerValSerGlySerProGlyGln
151015
SerIleThrIleSerCysThrGlyThrSerSerAspValGlyGlyTyr
202530
AsnSerValSerTrpTyrGlnGlnHisProGlyLysAlaProLysLeu
354045
MetIleTyrGluValSerAsnArgProSerGlyValSerAsnArgPhe
505560
SerGlySerLysSerGlyAsnThrAlaSerLeuThrIleSerGlyLeu
65707580
GlnAlaGluAspGluAlaAspTyrTyrCysAsnSerTyrThrSerThr
859095
SerMetValPheGlyGlyGlyThrLysLeuThrValLeu
100105
<210>23
<211>118
<212>PRT
<213>人工序列
<220>
<223>AliroVH
<400>23
GluValGlnLeuValGluSerGlyGlyGlyLeuValGlnProGlyGly
151015
SerLeuArgLeuSerCysAlaAlaSerGlyPheThrPheAsnAsnTyr
202530
AlaMetAsnTrpValArgGlnAlaProGlyLysGlyLeuAspTrpVal
354045
SerThrIleSerGlySerGlyGlyThrThrAsnTyrAlaAspSerVal
505560
LysGlyArgPheIleIleSerArgAspSerSerLysHisThrLeuTyr
65707580
LeuGlnMetAsnSerLeuArgAlaGluAspThrAlaValTyrTyrCys
859095
AlaLysAspSerAsnTrpGlyAsnPheAspLeuTrpGlyArgGlyThr
100105110
LeuValThrValSerSer
115
<210>24
<211>113
<212>PRT
<213>人工序列
<220>
<223>AliroVK
<400>24
AspIleValMetThrGlnSerProAspSerLeuAlaValSerLeuGly
151015
GluArgAlaThrIleAsnCysLysSerSerGlnSerValLeuTyrArg
202530
SerAsnAsnArgAsnPheLeuGlyTrpTyrGlnGlnLysProGlyGln
354045
ProProAsnLeuLeuIleTyrTrpAlaSerThrArgGluSerGlyVal
505560
ProAspArgPheSerGlySerGlySerGlyThrAspPheThrLeuThr
65707580
IleSerSerLeuGlnAlaGluAspValAlaValTyrTyrCysGlnGln
859095
TyrTyrThrThrProTyrThrPheGlyGlnGlyThrLysLeuGluIle
100105110
Lys