Coding

Part:BBa_K2520001

Designed by: Noa Eden, Dana Kadosh   Group: iGEM17_TECHNION-ISRAEL   (2017-10-25)


Single Chain Fragment Variable (scFv) of rat anti-murine IgM antibody

Single Chain Fragment Variable (scFv) A scFv is a fusion protein of the variable regions of an antibody (VH and VL) connected by a short linker peptide. This fusion protein retains the specificity of the original antibody despite the removal of the constant regions.

Figure 1: Structure of scFv. VH and VL are refer to variable heavy and variable light, respectively

This fusion protein can be secreted or displayed on the cellular membrane.

Results

Our scFv functions as anti-IgM. In order to verify that the anti-IgM construct is functional, we expressed it under two promoters- a mutant EF1a promoter BBa_K2520023 and CMV promoter and we tested the functionality using soluble murine IgM conjugated to a fluorophore.

Figure 2: fold change of the weighted average after extracellular staining with IgM conjugated to FITC, compared with non-transfected cells.


References

Lee, Timothy K., et al. "Production of engineered IgM-binding single-chain antibodies in Escherichia coli." Journal of Industrial Microbiology & Biotechnology 14.5 (1995): 371-376. Sequence and Features


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]


[edit]
Categories
//collections/immune_regulation/antibodies
Parameters
None