Abstract
Therapeutic proteins have recently received increasing attention because of their clinical potential. Currently, most therapeutic proteins are produced on a large scale using various cell culture systems. However, storing and transporting these therapeutic proteins at low temperatures makes their distribution expensive and problematic, especially for applications in remote locations. To this end, an emerging solution is to use point-of-care technologies that enable immediate and accessible protein production at or near the patient’s bedside. Here we present the development of “Therapeutics-On-a-Chip (TOC)”, an integrated microfluidic platform that enables point-of-care synthesis and purification of therapeutic proteins. We used fresh and lyophilized materials for cell-free synthesis of therapeutic proteins on microfluidic chips and applied immunoprecipitation for highly efficient, on-chip protein purification. We first demonstrated this approach by expressing and purifying a reporter protein, green fluorescent protein. Next, we used TOC to produce cecropin B, an antimicrobial peptide that is widely used to control biofilm-associated diseases. We successfully synthesized and purified cecropin B at 63 ng/μl within 6 h with a 92% purity, followed by confirming its antimicrobial functionality using a growth inhibition assay. Our TOC technology provides a new platform for point-of-care production of therapeutic proteins at a clinically relevant quantity.
Highlights
The ability to synthesize proteins in a point-of-care setting is an important step for reducing costs associated with storage and transportation of therapeutic proteins and enabling their widespread application in resourcepoor regions[1]
The majority of therapeutic proteins are produced on a large scale using various cell culture systems, including recombinant Escherichia coli[3] or yeast[4], mammalian[5], and plant[6] cells
We developed a highly integrated TOC system that enabled rapid and point-of-care synthesis and purification of therapeutic proteins based on a cell-free protein synthesis (CFPS) process
Summary
The ability to synthesize proteins in a point-of-care setting is an important step for reducing costs associated with storage and transportation of therapeutic proteins and enabling their widespread application in resourcepoor regions[1]. The majority of therapeutic proteins are produced on a large scale using various cell culture systems, including recombinant Escherichia coli[3] or yeast[4], mammalian[5], and plant[6] cells. These proteins are globally distributed from centralized foundries[7]. The capital cost of such a system will make it an unlikely solution for the developing world where the need is more for speedy production of different reagents for different diseases over producing mass quantities that require storage
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