Part:BBa_K1933002

anti-Norovirus GII.4 scFv

This part codes for anti-Norovirus GII.4 scFv (single-chain variable fragment) which derives from human 12A2 antibody. This scFv is suggested to prevent binding of human Norovirus to Hist-Blood Group Antigens[1].
This coding sequence was fused to INPNC-His (BBa_K1933001) or BclA-His (BBa_K1933000) in our project.
For more information, please visit [http://2016.igem.org/Team:Kyoto our wiki]

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal XhoI site found at 387
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Usage and Biology

NoV binding to Histo-blood group antigen (HBGA), expressed on the mucosal epithelium of the digestive tract and/or commensal bacteria, is suggested to be an important part of NoV invasion[2][3]. Also, NoV binding to 12A2 antibody in its HBGA binding site has been demonstrated previously[4]. The scFv coded in our part derives from antibody 12A2, so it also sterically blocks the binding of NoV to HBGA in theory, which might neutralize NoV using the same mechanism employed by human immune system.

We used this part for construction of ☆BBa_K1933200 and BBa_K1933201.

Characterization

Please see ☆BBa_K1933200 and BBa_K1933201 for full characterization.

MIT_MAHE 2020

Summary

Norovirus infections commonly lead to outbreaks of acute gastroenteritis and spread quickly, resulting in many health and economic challenges prior to diagnosis. This part codes for anti-Norovirus GII.4 scFv (single-chain variable fragment) which derives from human 12A2 antibody. This scFv is suggested to prevent binding of human Norovirus to Hist-Blood Group Antigen.

Reference

[1]Higo‐Moriguchi, Kyoko, et al. "Isolation of cross‐reactive human monoclonal antibodies that prevent binding of human Noroviruses to histo‐blood group antigens." Journal of medical virology 86.4 (2014): 558-567.

[2]Schroten, Horst, Franz-Georg Hanisch, and Grant S. Hansman. "Human Norovirus interactions with histo-blood group antigens and human milk oligosaccharides." Journal of virology 90.13 (2016): 5855-5859.

[3]Karst, Stephanie M., and Christiane E. Wobus. "A working model of how noroviruses infect the intestine." PLoS Pathog 11.2 (2015): e1004626.

[4]Shanker, Sreejesh, et al. "Structural basis for Norovirus neutralization by an HBGA blocking human IgA antibody." Proceedings of the National Academy of Sciences 113.40 (2016): E5830-E5837.

[5] Hurwitz AM, Huang W, Kou B, Estes MK, Atmar RL, Palzkill T (2017) Identification and Characterization of Single-Chain Antibodies that Specifically Bind GI Noroviruses. PLoS ONE 12(1): e0170162. https://doi.org/10.1371/journal.pone.0170162