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Publications (6 of 6) Show all publications
Björk, S. & Jönsson, H. (2019). Microfluidics for cell factory and bioprocess development. Current Opinion in Biotechnology, 55, 95-102
Open this publication in new window or tab >>Microfluidics for cell factory and bioprocess development
2019 (English)In: Current Opinion in Biotechnology, ISSN 0958-1669, E-ISSN 1879-0429, Vol. 55, p. 95-102Article in journal (Refereed) Published
Abstract [en]

Bioindustry is expanding to an increasing variety of food, chemical and pharmaceutical products, each requiring rapid development of a dedicated cell factory and bioprocess. Microfluidic tools are, together with tools from synthetic biology and metabolic modeling, being employed in cell factory and bioprocess development to speed up development and address new products. Recent examples of microfluidics for bioprocess development range from integrated devices for DNA assembly and transformation, to high throughput screening of cell factory libraries, and micron scale bioreactors for process optimization. These improvements act to improve the biotechnological engineering cycle with tools for building, testing and evaluating cell factories and bioprocesses by increasing throughput, parallelization and automation.

Place, publisher, year, edition, pages
Elsevier, 2019
Keywords
Cells, Cytology, Drug products, Microfluidics, Optimization, Bioprocess development, DNA assemblies, High throughput screening, Integrated device, Metabolic modeling, Parallelizations, Pharmaceutical products, Synthetic biology, Cell engineering, bioprocess, biotechnological procedures, cell factory, cell growth, cell population, cell selection, cell separation, priority journal, process optimization, Review, single cell analysis
National Category
Industrial Biotechnology
Identifiers
urn:nbn:se:kth:diva-236324 (URN)10.1016/j.copbio.2018.08.011 (DOI)000459949400015 ()2-s2.0-85053411311 (Scopus ID)
Funder
Novo NordiskKnut and Alice Wallenberg FoundationSwedish Research Council Formas
Note

QC 20181120

Available from: 2018-11-20 Created: 2018-11-20 Last updated: 2019-09-30
Wang, G., Björk, S., Huang, M., Liu, Q., Campbell, K., Nielsen, J., . . . Petranovic, D. (2019). RNAi expression tuning, microfluidic screening, and genome recombineering for improved protein production in Saccharomyces cerevisiae. Proceedings of the National Academy of Sciences of the United States of America, 116(19), 9324-9332
Open this publication in new window or tab >>RNAi expression tuning, microfluidic screening, and genome recombineering for improved protein production in Saccharomyces cerevisiae
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2019 (English)In: Proceedings of the National Academy of Sciences of the United States of America, ISSN 0027-8424, E-ISSN 1091-6490, Vol. 116, no 19, p. 9324-9332Article in journal (Refereed) Published
Abstract [en]

The cellular machinery that supports protein synthesis and secretion lies at the foundation of cell factory-centered protein production. Due to the complexity of such cellular machinery, the challenge in generating a superior cell factory is to fully exploit the production potential by finding beneficial targets for optimized strains, which ideally could be used for improved secretion of other proteins. We focused on an approach in the yeast Saccharomyces cerevisiae that allows for attenuation of gene expression, using RNAi combined with high-throughput microfluidic single-cell screening for cells with improved protein secretion. Using direct experimental validation or enrichment analysis-assisted characterization of systematically introduced RNAi perturbations, we could identify targets that improve protein secretion. We found that genes with functions in cellular metabolism (YDC1, AAD4, ADE8, and SDH1), protein modification and degradation (VPS73, KTR2, CNL1, and SSA1), and cell cycle (CDC39), can all impact recombinant protein production when expressed at differentially down regulated levels. By establishing a workflow that incorporates Cas9-mediated recombineering, we demonstrated how we could tune the expression of the identified gene targets for further improved protein production for specific proteins. Our findings offer a high throughput and semirational platform design, which will improve not only the production of a desired protein but even more importantly, shed additional light on connections between protein production and other cellular processes.

Place, publisher, year, edition, pages
National Academy of Sciences, 2019
National Category
Industrial Biotechnology Biochemistry and Molecular Biology Other Industrial Biotechnology
Identifiers
urn:nbn:se:kth:diva-250438 (URN)10.1073/pnas.1820561116 (DOI)000467226400031 ()2-s2.0-85065621578 (Scopus ID)
Note

QC 20190529

Available from: 2019-04-29 Created: 2019-04-29 Last updated: 2019-11-13Bibliographically approved
Björk, S. M., Sjostrom, S. L., Andersson-Svahn, H. & Jönsson, H. N. (2015). Metabolite profiling of microfluidic cell culture conditions for droplet based screening. Biomicrofluidics, 9(4), Article ID 044128.
Open this publication in new window or tab >>Metabolite profiling of microfluidic cell culture conditions for droplet based screening
2015 (English)In: Biomicrofluidics, ISSN 1932-1058, E-ISSN 1932-1058, Vol. 9, no 4, article id 044128Article in journal (Refereed) Published
Abstract [en]

We investigate the impact of droplet culture conditions on cell metabolic state by determining key metabolite concentrations in S. cerevisiae cultures in different microfluidic droplet culture formats. Control of culture conditions is critical for single cell/clone screening in droplets, such as directed evolution of yeast, as cell metabolic state directly affects production yields from cell factories. Here, we analyze glucose, pyruvate, ethanol, and glycerol, central metabolites in yeast glucose dissimilation to establish culture formats for screening of respiring as well as fermenting yeast. Metabolite profiling provides a more nuanced estimate of cell state compared to proliferation studies alone. We show that the choice of droplet incubation format impacts cell proliferation and metabolite production. The standard syringe incubation of droplets exhibited metabolite profiles similar to oxygen limited cultures, whereas the metabolite profiles of cells cultured in the alternative wide tube droplet incubation format resemble those from aerobic culture. Furthermore, we demonstrate retained droplet stability and size in the new better oxygenated droplet incubation format.

Keywords
Biomolecules;Cell culture;Cell proliferation;Cells;Dimensional stability;Glucose;Metabolism;Metabolites;Microfluidics;Yeast
National Category
Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:kth:diva-173782 (URN)10.1063/1.4929520 (DOI)000360311900030 ()2-s2.0-84940909670 (Scopus ID)
Funder
Science for Life Laboratory - a national resource center for high-throughput molecular bioscience
Note

QC 20150921

QC 20191008

Available from: 2015-09-21 Created: 2015-09-18 Last updated: 2019-10-08Bibliographically approved
Björk, S., Sjöström, S. L., Andersson Svahn, H. & Jönsson, H. N. Controlling cell metabolic state in droplet microfluidics.
Open this publication in new window or tab >>Controlling cell metabolic state in droplet microfluidics
(English)Manuscript (preprint) (Other academic)
National Category
Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:kth:diva-159308 (URN)
Note

QS 2015

Available from: 2015-01-29 Created: 2015-01-29 Last updated: 2015-01-29Bibliographically approved
Björk, S., Schappert, M. & Jönsson, H.Droplet microfluidic microcolony analysis of triacylglycerol yields in S. cerevisiae for high throughput screening.
Open this publication in new window or tab >>Droplet microfluidic microcolony analysis of triacylglycerol yields in S. cerevisiae for high throughput screening
(English)Manuscript (preprint) (Other academic)
National Category
Engineering and Technology
Identifiers
urn:nbn:se:kth:diva-259488 (URN)
Available from: 2019-09-16 Created: 2019-09-16 Last updated: 2019-10-18Bibliographically approved
Björk, S., Shabestary, K., Yao, L., Ljungqvist, E., Jönsson, H. & Hudson, E. P.Droplet microfluidic screening of a Synechocystis sp. CRISPRi library based on L-lactate production.
Open this publication in new window or tab >>Droplet microfluidic screening of a Synechocystis sp. CRISPRi library based on L-lactate production
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(English)Manuscript (preprint) (Other academic)
National Category
Engineering and Technology
Identifiers
urn:nbn:se:kth:diva-259487 (URN)
Note

QC 20191011

Available from: 2019-09-16 Created: 2019-09-16 Last updated: 2019-10-11Bibliographically approved
Organisations
Identifiers
ORCID iD: ORCID iD iconorcid.org/0000-0002-3722-5970

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