Part:BBa_K3056000
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).
//cds/reporter
//chassis/prokaryote/ecoli
//function/reporter/fluorescence
chassis | E. coli Rosetta-gami2 |
color | Yellow-green |
control | T7 (IPTG) |
emit | 517 nm |
excite | 506 nm |
target | delta-9-tetrahydrocannabinol (THC) |