Part:BBa_K3346003
Usage and Biology
Antibodies have great potential for the design of diagnostics (immunoassays) and therapeutic applications (immunotherapy) [1]. With such high affinity to their target molecules, antibodies can effectively capture target analytes in in vitro samples or in in vivo systems. However, the use of immunoassays and immunotherapy is limited in part because of their high cost [2]. While mass production can help reduce the expenses associated with the use of antibodies in a project design, the cost burden can make it hard for teams with limited resources to design and implement synthetic biology projects that use these molecules. We decided to use synthetic biology to produce the antibodies for immunoassays and immunotherapies to lower the cost of our diagnostic product and make it more accessible to clinics and laboratories across the globe. This could additionally help other teams planning to use antibodies in diagnostic or therapeutic applications by creating a family of parts that can be efficiently used with their desired sequences, thus creating a type of “plug and play” method for antibody production in E. coli. This BioBrick was designed for efficient use in the modified E. coli strain SHuffle, which has an oxidative cytoplasm and additional chaperone proteins to ensure the proper formation of disulfide bonds [3].
This composite part consists of the full length IgG of the therapeutic monoclonal antibody Silutuximab, which consists of the heavy chain, light chain and constant chain regions. The VH chain sequence is available in BBa_K3346000, the light chain sequence is available in BBa_K3346001, and the constant chain sequence is available in BBa_K3346002. An additional ribosomal binding site (BBa_ J61100) was placed between the light and heavy chains to ensure efficiency of translation [3]. This composite part was assembled using the RFC23 prefixes and suffixes to allow for proper in-frame assembly (six base pair scar). This composite part demonstrates the capability of our constant chain BioBrick to be combined with a desired variable chain sequence. This can be inserted into an expression plasmid with a strong T7 promoter and ribosomal binding site (BBa_J61100).
[1] Khan, F. H. (2014). Chapter 25 - Antibodies and Their Applications (A. S. Verma & A. B. T.-A. B. Singh (eds.); pp. 473–490). Academic Press. https://doi.org/https://doi.org/10.1016/B978-0-12-416002-6.00025-0
[2] Hernandez, I., Bott, S. W., Patel, A. S., Wolf, C. G., Hospodar, A. R., Sampathkumar, S., & Shrank, W. H. (2018). Pricing of monoclonal antibody therapies: higher if used for cancer? The American Journal of Managed Care, 24(2), 109–112. http://www.ncbi.nlm.nih.gov/pubmed/29461857
[3] Robinson, M.-P., Ke, N., Lobstein, J., Peterson, C., Szkodny, A., Mansell, T. J., Tuckey, C., Riggs, P. D., Colussi, P. A., Noren, C. J., Taron, C. H., DeLisa, M. P., & Berkmen, M. (2015). Efficient expression of full-length antibodies in the cytoplasm of engineered bacteria. Nature Communications, 6(1), 8072. https://doi.org/10.1038/ncomms9072
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