Research Projects

Synthetic biology has the potential to revolutionize societal health by producing personalized medicine through biomanufacturing recombinantly expressed biologics. However, current centralized facilities require high capital investment and lack modularity for small-scale manufacturing. Cell-Free Protein Synthesis (CFPS) is a powerful synthesis technology for in vitro expression and on-demand production of biologics. CFPS systems have been used to manufacture an antimicrobial biologic (cecropin B) with over 90% purity within 5 hours. However, the purification schemes of these biologics resemble those of centralized facilities, involving expensive affinity resins, destabilization during elution, and additional time for dialysis. A low-cost microfluidic platform is proposed that uses light as an external stimulus to purify recombinantly expressed biologics for localized and on-demand manufacturing. The foundational principle is the development of fusion constructs of a photocleavable protein (PhoCl) with the protein of interest (POI) with an N-terminal silica binding peptide (SBP) affinity tag. SBP is used due to its affinity towards inexpensive silica (glass), which will be the material of choice to fabricate the microfluidic platform. The working principle of the proposed technology is as follows: (1) CFPS of SBP-PhoCl-POI, (2) Loading of cell-free synthesized fusion proteins on the microfluidic glass platform, (3) Washing of non-specifically adsorbed proteins, (4) Light exposure to release the POI from PhoCl through photocleavage, and (5) Elution with the appropriate buffer as required. The proposed technology overcomes the challenges associated with existing centralized and microfluidic biomanufacturing systems, providing a low-cost, on-demand, and localized platform for producing personalized medicine.