2022-10-14
Nanobody Expression in WK6 E. coli and Extraction
Nanobody Expression in WK6 E. coli
Inoculation: Antibiotic, glucose, and magnesium were added to TB medium in baffled flasks. Inoculate until it reaches the desired cell density.
Induction: IPTG (Isopropyl β-D-1-thiogalactopyranoside) was added to induce nanobody expression.
Protein Extraction from Periplasm
Harvesting cells: After overnight incubation, bacterial cells were collected by centrifugation in small batches, removing the liquid supernatant and keeping the cell pellet.
Resuspension: The cell pellet was resuspended in a solution designed to maintain cell integrity while preparing for osmotic shock.
Osmotic Shock: to disrupt the periplasmic membrane and release the nanobodies
Final Centrifugation of the suspension to remove any debris
Periplasmic extract (containing the nanobodies) from the supernatant was collected.
The content and copyright of the experimental protocol belong to Ghent University.
Under no circumstances should any information be used for commercial purposes.
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