Difference between revisions of "Part:BBa K3573000:Design"

(→‎Design Notes)
(→‎Design Notes)
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2VH5 Heavy + Light Chains:
 
2VH5 Heavy + Light Chains:
−
EVQLLESGGGLVQPGGSLRLSAAASGFTFSTFSMNWVRQAPGKGLEWVSYISRTSKTIYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYVARGRFFDYWGQGTLVTVS
+
EVQLLESGGGLVQPGGSLRLSAAASGFTFSTFSMNWVRQAPGKGLEWVSYISRTSKTIYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYVARGRFFDY
−
IQMTQSPSSLSASVGDRVTITVRASQSISSYLNWYQQKPGEAPKLLIYSASVLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYAQQSVMIPMTFGQGTKVE
+
WGQGTLVTVSIQMTQSPSSLSASVGDRVTITVRASQSISSYLNWYQQKPGEAPKLLIYSASVLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYAQQSVMIP
 +
MTFGQGTKVE
 
​
 
​
 +
 
scFv(F8) with CPP and Linker:
 
scFv(F8) with CPP and Linker:
 
MTYTRRRFRRRRHRPRS  
 
MTYTRRRFRRRRHRPRS  
−
QVQLQESGGDLVQPGGSLKLSCAASGFTFSSYGMSWVRQTPDKRLELVATINSNGGSTFYPDSVKGRFTISRDNAKNTLYLQMSSLKSEDTAMYYCARRRNYPYYYGSRGTFDYWGQGTTVTVSS
+
QVQLQESGGDLVQPGGSLKLSCAASGFTFSSYGMSWVRQTPDKRLELVATINSNGGSTFYPDSVKGRFTISRDNAKNTLYLQMSSLKSEDTAMYYCARRRNYPY
 +
YYGSRGTFDYWGQGTTVTVSS 
 
GGGGSGGGGSGGGGS  
 
GGGGSGGGGSGGGGS  
−
DIELTQSPASLAVSLGQRATISCRASESVDSYGNSFMHWYQQKPGQPPKLLIYRALNLESGIPARFSGSGSRTDFTLTINPVEADDVATYYCQQSNEDPWTFGGGTKLEIKR
+
DIELTQSPASLAVSLGQRATISCRASESVDSYGNSFMHWYQQKPGQPPKLLIYRALNLESGIPARFSGSGSRTDFTLTINPVEADDVATYYCQQSNEDPWTFGG
 +
GTKLEIKR
 
​
 
​
 +
 
scFv(F8) with CPP and Linker, 2VH5's antigen binding residues grafted:
 
scFv(F8) with CPP and Linker, 2VH5's antigen binding residues grafted:
 
MTYTRRRFRRRRHRPRS
 
MTYTRRRFRRRRHRPRS
−
QVQLQESGGDLVQPGGSLKLSCAASGFTFSSYGMSWVRQTPDKRLELVATINRTGKTTYYPDSVKGRFTISRDNAKNTLYLQMSSLKSEDTAMYYCARGRFFDYWGQGTTVTVSS
+
QVQLQESGGDLVQPGGSLKLSCAASGFTFSSYGMSWVRQTPDKRLELVATINRTGKTTYYPDSVKGRFTISRDNAKNTLYLQMSSLKSEDTAMYYCARGRFFDY
 +
WGQGTTVTVSS
 
GGGGSGGGGSGGGGS
 
GGGGSGGGGSGGGGS
−
DIELTQSPASLAVSLGQRATISCRASESVDSYMHWYQQKPGQPPKLLIYRALNLESGIPARFSGSGSRTDFTLTINPVEADDVATYYCQQSVEDPWTFGGGTKLEIKR
+
DIELTQSPASLAVSLGQRATISCRASESVDSYMHWYQQKPGQPPKLLIYRALNLESGIPARFSGSGSRTDFTLTINPVEADDVATYYCQQSVEDPWTFGGGTKL
 +
EIKR
  
 
===Source===
 
===Source===

Revision as of 04:12, 23 October 2020


Single chain variable fragment (scFV) recognizing RAS


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This year's project is a continuation of 2019's project. Last year our team (Korea_HS) designed CPP-scFv(P5) by engineering scFv(F8) that is inherently hyperstable. We have shown that CPP-scFv(P5) can enter the cell and recognize its cognate target protein, lysozyme in a reducing environment.

In 2020, we have designed hyperstable, cell-penetrating scFv targeting RAS protein (scFv(RAS)) based on the structure of RAS: anti-RAS antibody (PDB ID: 2vh5)

2VH5 Heavy + Light Chains: EVQLLESGGGLVQPGGSLRLSAAASGFTFSTFSMNWVRQAPGKGLEWVSYISRTSKTIYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYVARGRFFDY WGQGTLVTVSIQMTQSPSSLSASVGDRVTITVRASQSISSYLNWYQQKPGEAPKLLIYSASVLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYAQQSVMIP MTFGQGTKVE ​

scFv(F8) with CPP and Linker: MTYTRRRFRRRRHRPRS QVQLQESGGDLVQPGGSLKLSCAASGFTFSSYGMSWVRQTPDKRLELVATINSNGGSTFYPDSVKGRFTISRDNAKNTLYLQMSSLKSEDTAMYYCARRRNYPY YYGSRGTFDYWGQGTTVTVSS GGGGSGGGGSGGGGS DIELTQSPASLAVSLGQRATISCRASESVDSYGNSFMHWYQQKPGQPPKLLIYRALNLESGIPARFSGSGSRTDFTLTINPVEADDVATYYCQQSNEDPWTFGG GTKLEIKR ​

scFv(F8) with CPP and Linker, 2VH5's antigen binding residues grafted: MTYTRRRFRRRRHRPRS QVQLQESGGDLVQPGGSLKLSCAASGFTFSSYGMSWVRQTPDKRLELVATINRTGKTTYYPDSVKGRFTISRDNAKNTLYLQMSSLKSEDTAMYYCARGRFFDY WGQGTTVTVSS GGGGSGGGGSGGGGS DIELTQSPASLAVSLGQRATISCRASESVDSYMHWYQQKPGQPPKLLIYRALNLESGIPARFSGSGSRTDFTLTINPVEADDVATYYCQQSVEDPWTFGGGTKL EIKR

Source

This DNA was synthesized

References