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3D-Printable Cellular Composites for the Production of Recombinant Proteins

Sim, Seunghyun and Hui, Yue and Tirrell, David A. (2022) 3D-Printable Cellular Composites for the Production of Recombinant Proteins. Biomacromolecules, 23 (11). pp. 4687-4695. ISSN 1525-7797. doi:10.1021/acs.biomac.2c00915.

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The incorporation of living cells into materials promises both significant challenges and new possibilities. Although recent years have seen important advances in this field, there is still much to be learned about engineering interfaces between cells and materials. Here, we present a new class of 3D-printable materials, based on poly(N-hydroxymethylacrylamide) (PNHMAA), in which the spore-forming bacterium Bacillus subtilis is effectively cross-linked into the surrounding polymeric scaffold. After dehydration and subsequent re-swelling in nutrient-rich media, embedded cells and spores become metabolically active and are capable of heterologous protein production and secretion. Strikingly, the leak-free scaffold allows protein production while preventing escape of embedded cells. The successful construction of complex three-dimensional structures by stereolithographic printing of living PNHMAA composite materials suggests utility in a broad range of applications.

Item Type:Article
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URLURL TypeDescription
Sim, Seunghyun0000-0002-4232-5917
Tirrell, David A.0000-0003-3175-4596
Additional Information:S.S. and Y.H. contributed equally to this work. S.S. thanks the Helen Hay Whitney Foundation for a Postdoctoral Fellowship. This work was supported by Defense Advanced Research Projects Agency Engineered Living Materials Agreement HR0011-17-2-0037 and by the Institute for Collaborative Biotechnologies through cooperative agreement W911NF-19-2-0026 from the U.S. Army Research Office. The content of the information does not necessarily reflect the position or the policy of the Government, and no official endorsement should be inferred. The authors thank Chiara Daraio for granting extensive access to the stereolithographic 3D printer in her laboratory, Eleftheria Roumeli for her help with the stereolithographic 3D printer, Julia Kornfield for use of the rheometer in her laboratory, and Red Lhota for discussion. Confocal imaging was performed in the Biological Imaging Facility, with the support of the Caltech Beckman Institute and the Arnold and Mabel Beckman Foundation. Scanning electron microscopy was performed at the Caltech Division of Geological and Planetary Sciences Analytical Facility, and cryo-electron microscopy was performed in the Beckman Institute Resource Center for Transmission Electron Microscopy at Caltech.
Funding AgencyGrant Number
Helen Hay Whitney FoundationUNSPECIFIED
Defense Advanced Research Projects Agency (DARPA)HR0011-17-2-0037
Army Research Office (ARO)W911NF-19-2-0026
Arnold and Mabel Beckman FoundationUNSPECIFIED
Caltech Beckman InstituteUNSPECIFIED
Issue or Number:11
Record Number:CaltechAUTHORS:20230125-514893900.19
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:118939
Deposited By: Research Services Depository
Deposited On:22 Feb 2023 19:27
Last Modified:22 Feb 2023 19:27

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