Skip to main content

Thank you for visiting You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Point-of-care production of therapeutic proteins of good-manufacturing-practice quality


Manufacturing technologies for biologics rely on large, centralized, good-manufacturing-practice (GMP) production facilities and on a cumbersome product-distribution network. Here, we report the development of an automated and portable medicines-on-demand device that enables consistent, small-scale GMP manufacturing of therapeutic-grade biologics on a timescale of hours. The device couples the in vitro translation of target proteins from ribosomal DNA, using extracts from reconstituted lyophilized Chinese hamster ovary cells, with the continuous purification of the proteins. We used the device to reproducibly manufacture His-tagged granulocyte-colony stimulating factor, erythropoietin, glucose-binding protein and diphtheria toxoid DT5. Medicines-on-demand technology may enable the rapid manufacturing of biologics at the point of care.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Get just this article for as long as you need it


Prices may be subject to local taxes which are calculated during checkout

Fig. 1: The current biologically derived medicines on demand, Bio-MOD, system (version 3.0), a miniature system for biologics manufacturing.
Fig. 2: Automated end-to-end expression and purification of G-CSF in Bio-MOD 2.0.
Fig. 3: Operational logic for Bio-MOD 3.0.
Fig. 4: Comparison of G-CSF-His produced in two identical Bio-MODs.
Fig. 5: G-CSF-His produced in the Bio-MOD.
Fig. 6: Real-time analytics from UV sensors.
Fig. 7: Characterization of GBP and EPO produced by Bio-MOD.


  1. Choi, E. J. & Ling, G. S. Battlefield medicine: paradigm shift for pharmaceuticals manufacturing. PDA J. Pharm. Sci. Technol. 68, 312 (2014).

    Article  PubMed  Google Scholar 

  2. Schellekens, H., Aldosari, M., Talsma, H. & Mastrobattista, E. Making individualized drugs a reality. Nat. Biotechnol. 35, 507–514 (2017).

    Article  PubMed  CAS  Google Scholar 

  3. Editorial. Patient-centered drug manufacture. Nat. Biotechnol. 35, 485 (2017).

    Article  CAS  Google Scholar 

  4. Adamo, A. et al. On-demand continuous-flow production of pharmaceuticals in a compact, reconfigurable system. Science 352, 61–67 (2016).

    Article  PubMed  CAS  Google Scholar 

  5. Smith, M. T., Wilding, K. M., Hunt, J. M., Bennett, A. M. & Bundy, B. C. The emerging age of cell-free synthetic biology. FEBS Lett. 588, 2755–2761 (2014).

    Article  PubMed  CAS  Google Scholar 

  6. Zemella, A., Thoring, L., Hoffmeister, C. & Kubick, S. Cell-free protein synthesis: pros and cons of prokaryotic and eukaryotic systems. ChemBioChem 16, 2420–2431 (2015).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  7. Tran, K. et al. Cell-free production of a therapeutic protein: expression, purification, and characterization of recombinant streptokinase using a CHO lysate. Biotechnol. Bioeng. 115, 92–102 (2018).

    Article  PubMed  CAS  Google Scholar 

  8. Martin, R. W. et al. Development of a CHO-based cell-free platform for synthesis of active monoclonal antibodies. ACS Synth. Biol. 21, 1370–1379 (2017).

    Article  CAS  Google Scholar 

  9. Boles, K. S. et al. Digital-to-biological converter for on-demand production of biologics. Nat. Biotechnol. 35, 672–675 (2017).

    Article  PubMed  CAS  Google Scholar 

  10. Pardee, K. et al. Portable, on-demand biomolecular manufacturing. Cell 167, 248–259 (2016).

    Article  PubMed  CAS  Google Scholar 

  11. Shukla, A. A., Hubbard, B., Tressel, T., Guhan, S. & Low, D. Downstream processing of monoclonal antibodies—application of platform approaches. Anal. Technol. Biomed. Life Sci. 15, 28–39 (2007).

    Article  CAS  Google Scholar 

  12. Hammerling, U., Kroon, R. & Sjödin, L. In vitro bioassay with enhanced sensitivity for human granulocyte colony-stimulating factor. J. Pharm. Biomed. Anal. 13, 9–20 (1995).

    Article  PubMed  CAS  Google Scholar 

  13. Brödel, A. K., Sonnabend, A. & Kubick, S. Cell-free protein expression based on extracts from CHO cells. Biotechnol. Bioeng. 111, 25–36 (2014).

    Article  PubMed  CAS  Google Scholar 

  14. Peñalber-Johnstone, C. et al. Optimizing cell-free protein expression in CHO: assessing small molecule mass transfer effects in various reactor configurations. Biotechnol. Bioeng. 114, 1478–1486 (2017).

    Article  PubMed  CAS  Google Scholar 

  15. Tiangco, C. et al. Measuring transdermal glucose levels in neonates by passive diffusion: an in vitro porcine skin model. Anal. Bioanal. Chem. 409, 3475–3482 (2017).

    Article  PubMed  CAS  Google Scholar 

  16. Collier, R. J. Multi-mutant diphtheria toxin vaccines. US patent 7115725 B2 (2006).

  17. Giannini, G., Rappuoli, R. & Ratti, G. The amino-acid sequence of two non-toxic mutants of diphtheria toxin: CRM45 and CRM197. Nucleic Acids Res. 12, 4063–4069 (1984).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  18. Leader, B., Baca, Q. J. & Golan, D. E. Protein therapeutics: a summary and pharmacological classification. Nat. Rev. Drug Discov. 7, 21–39 (2008).

    Article  PubMed  CAS  Google Scholar 

  19. Gurramkonda, C. et al. Improving the recombinant human erythropoietin glycosylation using microsome supplementation in CHO cell-free system. Biotechnol. Bioeng. 115, 1253–1264 (2018).

    Article  PubMed  CAS  Google Scholar 

  20. Rathore, A. S. & Winkle, H. Quality by design for biopharmaceuticals. Nat. Biotechnol. 27, 26–34 (2009).

    Article  PubMed  CAS  Google Scholar 

  21. Swartz, J. R. Transforming biochemical engineering with cell-free biology. AIChE J. 58, 5–13 (2012).

    Article  CAS  Google Scholar 

  22. Buntru, M., Vogel, S., Stoff, K., Spiegel, H. & Schillberg, S. A versatile coupled cell-free transcription-translation system based on tobacco BY-2 cell lysates. Biotechnol. Bioeng. 112, 867–78 (2015).

    Article  PubMed  CAS  Google Scholar 

  23. Burgenson, D. et al. Rapid recombinant protein expression in cell-free extracts from human blood. Sci. Rep. (2018).

  24. Ding, W., Madsen, G., Mahajan, E., O’Connor, S. & Wong, K. Standardized extractables testing protocol for single-use systems in biomanufacturing. Pharm. Eng. 34, 1–11 (2014).

    CAS  Google Scholar 

Download references


We thank DARPA Biologically-derived Medicines on Demand (Bio-MOD) Project Grant (N66001-13-C-4023, ‘CASTing Biologics on Demand’) for financial support. G. Ling (Retd. US Army Colonel and former DARPA Battlefield Medicine Program Director) conceived the vision for Bio-MOD based on his difficulties in securing medicines to treat patients during deployments in Iraq and Afghanistan. This paper is dedicated to him. We thank E. Choi, J. Lewin, A. Bryon, M. Zamisch, T. McQuade, B. Ringeisen, G. Kost, K. Pankratz, R. Cecil, B. Webb, A. Bose, B. Junker, W.-L. Ling, M. McGinnis, P. Latham and R. Gopinath for various discussions, encouragement and support, and I. Shaffer of the Molecular Characterization and Analysis Complex, University of Maryland Baltimore County for sample analysis of leachables and extractables. We thank E. Gutierrez for his assistance with the illustrations. S.V. acknowledges Y. Xia of the MNMR Structural Biology Centre, University of Minnesota for technical help in the NMR spectroscopy. We thank the FDA Emerging Technology Team for helpful guidance and discussions. Disclaimer: This work was conducted while S.V. was employed at the University of Maryland School of Pharmacy. The opinions expressed in the article are the author’s own and do not reflect the view of the National Institutes of Health, the Department of Health and Human Services, or the United States government. No endorsement of this work by the Food and Drug Administration, National Institutes of Health, the Department of Health and Human Services, DARPA, Department of Defense or the United States government is implied.

Author information

Authors and Affiliations



M.A., A.A., D.B., X.G., Y.K., M.P., B.P., G.R., D.T., M.T., K.Tr. and B.W. designed, assembled and tested the Bio-MOD system. R.A., A.M., K.M., C.G., C.P., M.P., B.P., D.T. and K.Tr. maintained and reviewed the batch records. D.F., Y.L., H.G., S.V., A.Z. and S.D. did the protein purification. M.C., C.G., P.J., M.P., K.V. and D.W. did the cloning. R.A., S.Bo., S.Br., S.D., X.G., H.G., Y.L., K.M., C.P., M.P., B.P., A.R., P.R., S.S., K.Ta., L.T., K.V., S.V., J.W. and W.L. did the product analysis including gels, activity assays, mass spectrometry, protein concentration, sterility, silver staining, lysate stability, leachables and extractables. S.Br., D.B., M.C., C.G., P.J., M.P., K.Tr., K.V. and S.V. did the protein expression. D.F., X.G., C.G., Y.K., A.M., G.R., L.T., S.V., D.W. and K.V. did the overall and subsystem experimental design, execution and analysis. D.F., X.G., Y.K., C.P., B.P., G.R., L.T., K.V., S.V. and D.W. analysed the data.

Corresponding author

Correspondence to Govind Rao.

Ethics declarations

Competing interests

G.R., Y.K., L.T., X.G. and D.F. are listed as inventors on the United States patents 9,388,373 ‘Microscale Bioprocessing System and Method for Protein Manufacturing’ and 9,982,227 ‘System and Method for Production of On-Demand Proteins in a Portable Unit for Point of Care Delivery’.

Additional information

Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Supplementary Information

Supplementary figures and tables.

Reporting Summary

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Adiga, R., Al-adhami, M., Andar, A. et al. Point-of-care production of therapeutic proteins of good-manufacturing-practice quality. Nat Biomed Eng 2, 675–686 (2018).

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI:


Quick links

Nature Briefing: Translational Research

Sign up for the Nature Briefing: Translational Research newsletter — top stories in biotechnology, drug discovery and pharma.

Get what matters in translational research, free to your inbox weekly. Sign up for Nature Briefing: Translational Research