Process analytical technology (PAT) is critical for the manufacture of high-quality biologics as it enables continuous, real-time and on-line/at-line monitoring during biomanufacturing processes. The conventional analytical tools currently used have many restrictions to realizing the PAT of current and future biomanufacturing. Here we describe a nanofluidic device for the continuous monitoring of biologics’ purity and bioactivity with high sensitivity, resolution and speed. Periodic and angled nanofilter arrays served as the molecular sieve structures to conduct a continuous size-based analysis of biologics. A multiparameter quality monitoring of three separate commercial biologic samples within 50 minutes has been demonstrated, with 20 µl of sample consumption, inclusive of dead volume in the reservoirs. Additionally, a proof-of-concept prototype system, which integrates an on-line sample-preparation system and the nanofluidic device, was demonstrated for at-line monitoring. Thus, the system is ideal for on-site monitoring, and the real-time quality assurance of biologics throughout the biomanufacturing processes.
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We thank P. Mao for providing advice on the nanofilter fabrication, the MIT Microsystems Technology Laboratories for support in the fabrication, P. W. Barone for discussion and assistance with the sample handling and J.-F. P. Hamel for his support in the cell culture and analysis. This work was mainly supported by the Defense Advanced Research Projects Agency and SPAWAR Systems Center Pacific (N66001-13-C-4025) to S.H.K., D.C., W.O., T.K., P.K. and J.H. a Siebel fellowship to W.O. and a Samsung Scholarship to T.K.
J.H. and S.H.K. have filed a patent application for the nanofilter device. P.K. and D.C. plan on filing patent applications for the peptide ligands.
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Ko, S., Chandra, D., Ouyang, W. et al. Nanofluidic device for continuous multiparameter quality assurance of biologics. Nature Nanotech 12, 804–812 (2017). https://doi.org/10.1038/nnano.2017.74
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