Difference between revisions of "Part:BBa K3056000"
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+ | __NOTOC__ | ||
+ | <partinfo>BBa_K3056000 short</partinfo> | ||
+ | '''α -THC Antibody Conjugate to mNG:''' | ||
+ | Validation of expression, fluorescence, and binding ability to Δ9-tetrahydrocannabinol (henceforth, THC). | ||
+ | <br/> | ||
+ | <br/> | ||
+ | '''Components''' | ||
+ | This part consists of a periplasmic localization signal (PLS), and an anti-Δ9-tetrahydrocannabinol ScFv linked to mNG at the C-terminal. Note that this part contains a stop codon. | ||
+ | <br/> | ||
+ | <br/> | ||
+ | '''Summary''' | ||
+ | A fluorescently labelled anti-THC antibody was successfully produced in E. coli. The antibody fragment was able to bind THC soaked lipophilic membranes, with a sensitivity of 0.1 mg/mL. Future experiments may aim to increase the sensitivity of the THC assay by changing the fluorescent tags or testing a larger library of lipophilic membranes. | ||
+ | <br/> | ||
+ | <br/> | ||
+ | '''Background''' | ||
+ | Here we designed a fluorescent anti-THC antibody, optimized for E. coli expression. Recombinant antibody expression in E. coli is notoriously challenging, as typical IgG proteins require post-translational modifications. However, Recombinant expression of antibodies can be made possible by truncating antibody fragment ('''Fig. 1'''). | ||
+ | [[File:T--Queens_Canada--Bronze_Antibody_Trunc.jpg|thumb|600px|left|<b>Figure 1.</b>igure 1. The modification of an IgG protein to a Fab and ScFv, which can be expressed in E. coli. In the case of the anti-THC antibody, the protein was conjugated to a fluorescent protein at the C-terminal.]] |
Revision as of 21:12, 10 October 2019
Anti-THC antibody fragment (ScFv) linked to mNG at the C-terminal
α -THC Antibody Conjugate to mNG:
Validation of expression, fluorescence, and binding ability to Δ9-tetrahydrocannabinol (henceforth, THC).
Components
This part consists of a periplasmic localization signal (PLS), and an anti-Δ9-tetrahydrocannabinol ScFv linked to mNG at the C-terminal. Note that this part contains a stop codon.
Summary
A fluorescently labelled anti-THC antibody was successfully produced in E. coli. The antibody fragment was able to bind THC soaked lipophilic membranes, with a sensitivity of 0.1 mg/mL. Future experiments may aim to increase the sensitivity of the THC assay by changing the fluorescent tags or testing a larger library of lipophilic membranes.
Background
Here we designed a fluorescent anti-THC antibody, optimized for E. coli expression. Recombinant antibody expression in E. coli is notoriously challenging, as typical IgG proteins require post-translational modifications. However, Recombinant expression of antibodies can be made possible by truncating antibody fragment (Fig. 1).