Coding

Part:BBa_K4477004

Designed by: Miranda Khoury   Group: iGEM22_Virginia   (2022-09-06)


McPC603 anti-oxLDL scFv antibody fragment

This sequence codes for a short chain variable fragment (scFv) derived from the anti-oxLDL McPC603 antibody (1). Ox-LDL stands for oxidized low density lipoprotein, and it is a biomarker of many inflammatory diseases such as atherosclerosis. Specifically, McPC603 targets a chemical group called oxidized phosphocholine found within the lipid core of oxLDL. The sequence codes for the following in sequential order: McPC603 variable heavy chain, a (G4S)3 linker sequence, and McPC603 variable light chain.

The variable heavy and light chain of McPC603 are connected by a (G4S)3 linker sequence (2). This linker sequence was chosen because it was characterized to enhance protein expression of recombinant antibodies. Note that the protein resulting from this sequence will only fold correctly in organisms capable of producing disulfide bonds, as disulfide bonds are needed for the proper folding of antibodies. Team Virginia 2022 used SHuffle E. coli as the expression chassis.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 355
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 661
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None