(496c) On-Chip Manufacturing of Synthetic Proteins for Point-of-Care Therapeutics

Authors: 
Sheng, J., Virginia Polytechnic Institute and State University
Murphy, T., Virginia Tech
Lu, C., Virginia Tech
Feng, X., Virginia Polytechnic Institute and State University
Therapeutic proteins have recently received increasing attention because of their clinical potential. The annual sales of natural and modified therapeutic proteins approved for clinical use in the European Union and the United States reached $108 billion in 2010. Currently, most therapeutic proteins are produced on a large scale using various cell culture systems. However, the requirement for continuous cold storage of therapeutic proteins makes them difficult in delivery, especially at remote locations. To this end, an emerging solution is point-of-care technologies that enable immediate and accessible health care at or near the patient's bedside. Here, we present the development of “Therapeutics-On-A-Chip,” an integrated microfluidic platform that enables point-of-care synthesis and purification of therapeutic proteins. This system has an integrated power supply, syringe pump, and microfluidic chips packaged in a portable box that is similar in size to a standard medical box. The microfluidic chips are disposable and could be used in the portable system during protein synthesis and discarded after use. In Therapeutics-On-A-Chip, we use lyophilized materials for cell-free synthesis of therapeutic proteins on microfluidic chips and applied affinity chromatography for highly efficient, on-chip protein purification. We first demonstrated this approach by expressing and purifying a reporter protein, green fluorescence protein. Then, we used Therapeutics-On-A-Chip to produce Cecropin B, an antimicrobial peptide that is widely used to control biofilm-associated diseases. We successfully synthesized and purified one dose of Cecropin B (~6 μg) within 4 hours, followed by confirming its antimicrobial functionality via a growth inhibition assay. We expect our Therapeutics-On-A-Chip technology will supply therapeutic proteins, whenever and wherever they are needed, to precisely control various diseases worldwide.