Optimising open enzyme purification using 3D-printing and automation

Local manufacturing and purification of proteins may overcome some hurdles for researchers in resource-constrained contexts; this project aims to identify purification methods that can later be scaled up to develop cost-optimised protocols for distributed production of research enzymes.

The Idea

The number of openly licensed and off-patent enzymes for molecular biology is growing rapidly. However, legal rights to reuse and redistribute enzymes are insufficient when practical routes for global access to the biological materials are not in place. Local manufacturing and purification of proteins may overcome some constraints and make reagents more affordable for researchers in resource-constrained contexts, from low income countries to community laboratories and small companies. Synthetic biology techniques combined with automation now allow rapid combinatorial testing of biological constructs which we will harness to select optimised pairings of off-patent binding domains and substrates for column or well-based enzyme purification.

Specifically, we will use a 96-well 3D-printed reaction chamber (designed by Clayton Rabideau and supported by the OpenPlant Fund in 2017) as a semi-automated combinatorial testing device for off-patent binding domains where an attached marker protein will be column-purified using a variety of affordable materials. The affinity, yield and purity data will then be openly published online. The most successful binding domain and substrate combinations will form the basis of a follow-on project applying the domains to a curated collection of off-patent enzymes for basic molecular and synthetic biology applications. This will allow development of lower-cost protocols for manufacture and distribution and assessment of the economic feasibility of distributed manufacturing of research enzymes.