Platformed Inteins: A linked orthogonal toolkit
With a clear demand in biologics production, there is no shortage of ideas for innovation however bioproduction is one of many limiting factors that exist for moving drugs into clinical trials due to their fine tuned engineering and target specificity requirements.
Biologics make up 25% of all FDA approved drugs from 1997-2021 (De la Torre & Albericio, 2022). The move from traditional to emerging therapies is thus closely linked with bioproduction and producing enough for preclinical testing.
Statistics demonstrate significant amounts of hazardous and non-hazardous waste generation in global pharmaceutical companies. While efforts to reduce general and chemical waste have helped, more is to be done to promote sustainability.
Current biomanufacturing platforms are not enough for sustained bioproduction to meet the demand.
PILOT aims to develop a modular, biosynthesis toolkit for improved in vitro protein production that tackles the following pillars:
To aid with our cell free platform development, we designed an aerated bioreactor to culture our strain of Vibrio natriegens for cell extract production.
Our protein modelling subteam additionally modelled the general stability and thermodynamic stability of our Proof of Concept.
To aid with our cell free platform development, we designed an aerated bioreactor to culture our strain of Vibrio natriegens cell extract production.
Protein Production Efficiency
Employing rapid dividing Vibrio natriegens as a chassis for cell free protein synthesis.
Energy Usage Efficiency
Using oxidative phosphorylation instead of substrate level through split inteins.
Protein Purification Efficiency
Thiol-sensitive intein for affinity tag purification aid.
Our end goal was to test this system in a range of protein families from reporter proteins to cytokines like IL-10 thus demonstrating the modularity of our system for use in biologics production.
Behind Our Name PILOT: Platformed Inteins: A linked orthogonal toolkit.
Scaling up intein technology as part of a high throughput modular platform for the purposes of linking biosynthesis and protein purification. These processes of biosynthesis and purification occur independently but simultaneously in the same cell lysate. Therein lies the orthogonality of our toolkit.