On-chip manufacturing of synthetic proteins for point-of-care therapeutics View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2019-12

AUTHORS

Travis W. Murphy, Jiayuan Sheng, Lynette B. Naler, Xueyang Feng, Chang Lu

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. A microfluidic reactor enables researchers to synthesize therapeutic proteins at the point of care. Therapeutic proteins are usually produced in a central location, requiring costly refrigeration and transport to the location where they’re administered. This hinders care for patients in remote or resource-scarce areas. Engineers from the United States’ Virginia Tech, led by Chang Lu, have now developed a microfluidic reactor that uses cell components and a DNA template, completed by a protein purification device. The team validated their device by producing an easily quantifiable protein GFP, before producing therapeutically viable concentrations of cecropin B, a protein used in the treatment of microbial infections. The functionality of the synthesized cecropin B was confirmed by its successful inhibition of Escherichia coli. The devices are low-cost, and future research could extend their utility to other clinically beneficial proteins. More... »

PAGES

13

Journal

TITLE

Microsystems & Nanoengineering

ISSUE

1

VOLUME

5

Author Affiliations

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/s41378-019-0051-8

DOI

http://dx.doi.org/10.1038/s41378-019-0051-8

DIMENSIONS

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41 schema:description 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. A microfluidic reactor enables researchers to synthesize therapeutic proteins at the point of care. Therapeutic proteins are usually produced in a central location, requiring costly refrigeration and transport to the location where they’re administered. This hinders care for patients in remote or resource-scarce areas. Engineers from the United States’ Virginia Tech, led by Chang Lu, have now developed a microfluidic reactor that uses cell components and a DNA template, completed by a protein purification device. The team validated their device by producing an easily quantifiable protein GFP, before producing therapeutically viable concentrations of cecropin B, a protein used in the treatment of microbial infections. The functionality of the synthesized cecropin B was confirmed by its successful inhibition of Escherichia coli. The devices are low-cost, and future research could extend their utility to other clinically beneficial proteins.
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