The Open Enzyme Manufacturing Masterclass was piloted in Ghana this February in collaboration with Kwame Nkrumah University of Science and Technology. 12 participants undertook a week of training and successfully produced OpenVent, an enzyme for amplifying DNA for research, education and diagnostics.
Over the course of a week the participants, facilitated by OBL Team members Harry Akligoh, Sam Witham, Dr Samuel Asamoah-Sakyi and KNUST colleague Selorm Segbefia, covered the basic principle of assembling DNA components, expressing recombinant proteins in bacteria and performing quality controls on the resulting enzymes. Daily activities included;
Participants were welcomed and introduced to the work of the Open Bioeconomy Lab. Following suit were knowledge modules and skills modules which happened interchangeably at defined times during the day (same for the remaining days). The knowledge modules for day one included; Bacteria as host for recombinant protein expression, Practical Primer Design and Using DNA (cleanup, gel extraction, ligation, restriction digestion, cloning). The skills module also included; PCR with Ben Taq Hot Start, Gel electrophoresis, Media preparation and gel extraction using gel extraction. Day one ended with lots of excitement for participants and anticipations for day two’s activities.
The format of Day two was not different from that of Day one. Participants were again engaged with knowledge and skills modules to build upon skills and knowledge acquired from Day one. The knowledge modules included; DNA Assembly, Introduction to Synthetic Biology and Horizontal Gene Transfer. Therefore, to demonstrate the practicality of these concepts participants had a laboratory session on Klenow assembly to demonstrate DNA assembly methods, competent cell preparation and cell transformation to demonstrate horizontal gene transfer which are key techniques underpinning synthetic biology.
Participants continued learning and building new skills leveraging what they were introduced to in the previous days. For day three, the knowledge modules the participants were taught included; Biosafety, Ethics and Intellectual Property Right (IPR) in Biotechnology and Protein expression. The main skill modules juxtaposed to the protein expression knowledge module to enhance participants understanding included media preparation and inoculation of media with BL21 (DE3) E.coli and overnight incubation for subsequent downstream processing.
Day four proved to be the tedious of all days with several laboratory sessions. Participants began day four in the lab. They started off by continuing with the downstream processing of the overnight culture media and prepared 10ml culture media from the overnight culture and incubated it for 2-3 hrs and monitored cell growth using McFarland Standard until cell growth reached exponential phase. In the same culture after 2-3 hrs participants added IPTG to induce protein expression and incubated for an additional 2 hrs. After 2 hrs of protein induction and expression, participants had their Deep Vent DNA polymerase expressed in BL21(DE3) E.coli cells as cellular reagents. Knowledge modules that characterized day four included; Analyzing and Characterizing proteins, Storage of protein in cells and Optimization of protein expression in Bioreactors.
Day five was the final day of the one week journey of introducing Ghanaian scientists and postgraduate students to Low-cost protein biomanufacturing. Sessions that climaxed day five included; a brainstorming and Q & A session, knowledge module on Open Enzyme collection and final laboratory practicals to confirm protein expression using SDS PAGE.
The one week masterclass was very hectic and fun, however, it was a refreshing and rewarding opportunity for us because it reminded us of our contribution toward enhancing access to basic research tools through local capacity building in resource limited environments.