Protein_Domain
3-24 ScFv

Part:BBa_K3767000

Designed by: Eric Wen & Griffin Watson-Boehnisch   Group: iGEM21_Queens_Canada   (2021-06-16)


3-24 Anti-OspA ScFv optimized for E. Coli

Description

An antibody in ScFv format, or single chain fragment variable format, consists of the heavy (VH) and light (VL) chains of an antibody joined together by a peptide linker [2]. The 3-24 ScFv was created to detect Borrelia Burgdorferi, the bacteria causing Lyme Disease, through the detection of outer surface protein A (OspA). The ScFv was designed based on previous research by Ghosh and Huber [1], which we optimized for increased binding affinity to OspA and E. coli expression.

Usage and Biology

ScFvs are the smallest unit of an antibody with antigen-binding functions, and can be easily altered to increase affinity and specificity for certain antigens. They do not require the post-translational modifications essential for the creation of full-size IgG antibodies, and can thus be easily expressed by common bacteria such as E. coli[3]. The 3-24 ScFv was designed based on research by Ghosh and Huber[1], and is used to detect Borrelia Burgdorferi through the detection of outer surface protein A (OspA). It was designed as a diabody, containing the heavy and light chain of the Fv region, complementarity-determining regions (CDRs) for OspA to bind to, and a (GGGS)3 peptide linker to attach both chains. A glycine linker was used because they are very flexible and limit strain between each protein chain.

Figure 1: The modification of an IgG protein into an scFv diabody, containing 1 heavy and 1 light chain attached with a peptide linker. The scFv presents multiple CDRs in its antigen binding site that can bind to OspA.



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]


References

1. Ghosh, S., and Huber, B. T. (2007) Clonal diversification in OspA-specific antibodies from peripheral circulation of a chronic Lyme arthritis patient. J. Immunol. Methods. 321, 121–134

2. Zuhaida Asra Ahmad, Swee Keong Yeap, Abdul Manaf Ali, Wan Yong Ho, Noorjahan Banu Mohamed Alitheen, Muhajir Hamid, "scFv Antibody: Principles and Clinical Application", Journal of Immunology Research, vol. 2012, Article ID 980250, 15 pages, 2012. https://doi.org/10.1155/2012/980250

3.Warkentin, R. (2019, September 8). Part:BBa_K3056000. Registry of Standard Biological Parts. iGEM Registry. https://parts.igem.org/wiki/index.php?title=Part%3ABBa_K3056000.

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