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

(→‎Design Notes)
(→‎Design Notes)
Line 8: Line 8:
 
===Design Notes===
 
===Design Notes===
  
−
This year's project is a continuation of 2019's project. Last year our team (Korea_HS) designed and tested CPP-scFv(P5) for its ability to enter the cell and recognize its cognate target protein, lysozyme in a reducing environment.
+
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)
 
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:
 +
EVQLLESGGGLVQPGGSLRLSAAASGFTFSTFSMNWVRQAPGKGLEWVSYISRTSKTIYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYVARGRFFDYWGQGTLVTVS
 +
IQMTQSPSSLSASVGDRVTITVRASQSISSYLNWYQQKPGEAPKLLIYSASVLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYAQQSVMIPMTFGQGTKVE
 +
​
 +
scFv(F8) with CPP and Linker:
 +
MTYTRRRFRRRRHRPRS
 +
QVQLQESGGDLVQPGGSLKLSCAASGFTFSSYGMSWVRQTPDKRLELVATINSNGGSTFYPDSVKGRFTISRDNAKNTLYLQMSSLKSEDTAMYYCARRRNYPYYYGSRGTFDYWGQGTTVTVSS
 +
GGGGSGGGGSGGGGS
 +
DIELTQSPASLAVSLGQRATISCRASESVDSYGNSFMHWYQQKPGQPPKLLIYRALNLESGIPARFSGSGSRTDFTLTINPVEADDVATYYCQQSNEDPWTFGGGTKLEIKR
 +
​
 +
scFv(F8) with CPP and Linker, 2VH5's antigen binding residues grafted:
 +
MTYTRRRFRRRRHRPRS
 +
QVQLQESGGDLVQPGGSLKLSCAASGFTFSSYGMSWVRQTPDKRLELVATINRTGKTTYYPDSVKGRFTISRDNAKNTLYLQMSSLKSEDTAMYYCARGRFFDYWGQGTTVTVSS
 +
GGGGSGGGGSGGGGS
 +
DIELTQSPASLAVSLGQRATISCRASESVDSYMHWYQQKPGQPPKLLIYRALNLESGIPARFSGSGSRTDFTLTINPVEADDVATYYCQQSVEDPWTFGGGTKLEIKR
  
 
===Source===
 
===Source===

Revision as of 04:10, 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: EVQLLESGGGLVQPGGSLRLSAAASGFTFSTFSMNWVRQAPGKGLEWVSYISRTSKTIYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYVARGRFFDYWGQGTLVTVS IQMTQSPSSLSASVGDRVTITVRASQSISSYLNWYQQKPGEAPKLLIYSASVLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYAQQSVMIPMTFGQGTKVE ​ scFv(F8) with CPP and Linker: MTYTRRRFRRRRHRPRS QVQLQESGGDLVQPGGSLKLSCAASGFTFSSYGMSWVRQTPDKRLELVATINSNGGSTFYPDSVKGRFTISRDNAKNTLYLQMSSLKSEDTAMYYCARRRNYPYYYGSRGTFDYWGQGTTVTVSS GGGGSGGGGSGGGGS DIELTQSPASLAVSLGQRATISCRASESVDSYGNSFMHWYQQKPGQPPKLLIYRALNLESGIPARFSGSGSRTDFTLTINPVEADDVATYYCQQSNEDPWTFGGGTKLEIKR ​ scFv(F8) with CPP and Linker, 2VH5's antigen binding residues grafted: MTYTRRRFRRRRHRPRS QVQLQESGGDLVQPGGSLKLSCAASGFTFSSYGMSWVRQTPDKRLELVATINRTGKTTYYPDSVKGRFTISRDNAKNTLYLQMSSLKSEDTAMYYCARGRFFDYWGQGTTVTVSS GGGGSGGGGSGGGGS DIELTQSPASLAVSLGQRATISCRASESVDSYMHWYQQKPGQPPKLLIYRALNLESGIPARFSGSGSRTDFTLTINPVEADDVATYYCQQSVEDPWTFGGGTKLEIKR

Source

This DNA was synthesized

References