Part:BBa_K2876007

anti-aflatoxinB1 ScFv1

This is a single-chain antibody targeting aflatoxin. We took the sequence from BBa_K2247006 and simply codon optimized it for E. coli rather than S. cerevisiae.

For our small molecule test case we chose to detect aflatoxin, a small molecule produced by Aspergillus fungi species that infects many agricultural crops, including cannabis.2,3 Aflatoxin also causes a number of health problems in people when inhaled or ingested by immunocompromised patients.7 We fused two single chain antibodies targeting aflatoxin (BBa_K2876005 and BBa_K2876003) to the lambda repressor and alpha subunit of RNA-polymerase III (BBa_K2876002) described in Dove, Joung, and Hochschild in order to create two fusion proteins capable of activating transcription in the presence of aflatoxin (Figure 4 small molecule). For our reporter we used the previously described pOL2-62 from Dove, Joung, and Hochschild fused to a RFP. See more information about our reporter below.

MIT_MAHE 2020

Summary

Aflatoxins are a group of secondary metabolites produced by Aspergillus flavus and A. parasiticus which are highly toxic. They contaminate a vast array of foods and agricultural commodities, particularly maize, groundnuts, oilseeds, and tree nuts in tropical and subtropical regions of the world.This part refers to the single-chain anti-Aflatoxin B1 antibody, conferring the binding affinity to aflatoxin B1. Aflatoxin B1 is a major type of aflatoxin contaminating our everyday foods, and is a strong carcinogen to humans.

References

Li, X., Li, P., Zhang, Q., Li, Y., Zhang, W., & Ding, X. (2012). Molecular characterization of monoclonal antibodies against aflatoxins: a possible explanation for the highest sensitivity. Analytical chemistry, 84(12), 5229–5235. https://doi.org/10.1021/ac202747u

Cheng, M., Chan, S. Y., Zhao, Q., Chan, E. Y., Au, S. W., Lee, S. S., & Cheung, W. T. (2011). Construction and characterization of single-chain variable fragment antibody library derived from germline rearranged immunoglobulin variable genes. PloS one, 6(11), e27406. https://doi.org/10.1371/journal.pone.0027406

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 526
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]