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Jönsson, Håkan, PhDORCID iD iconorcid.org/0000-0001-5232-0805
Alternative names
Publications (10 of 41) 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: 2019-11-26Bibliographically approved
Langer, K. & Jönsson, H. (2019). Rapid production and recovery of cell spheroids by automated droplet microfluidics.
Open this publication in new window or tab >>Rapid production and recovery of cell spheroids by automated droplet microfluidics
2019 (English)Manuscript (preprint) (Other academic)
Abstract [en]

Droplet microfluidics enables high throughput cell processing, analysis and screening by miniaturizing the reaction vessels to nano- or pico-liter water-in oil droplets, but like many other microfluidic formats, droplet microfluidics have not been interfaced with or automated by laboratory robotics. Here we demonstrate automation of droplet microfluidics based on an inexpensive liquid handling robot for the automated production of human scaffold-free cell spheroids, using pipette actuation and interfacing the pipetting tip with a droplet generating microfluidic chip. In this chip we produce highly mono-disperse 290μm droplets with diameter CV of 1.7%. By encapsulating cells in these droplets, we produce cell spheroids in droplets and recover them to standard formats at a throughput of 85000 spheroids per microfluidic circuit per hour. The viability of the cells in spheroids remains high after recovery only decreased by 4% starting from 96% after 16 hours incubation in nanoliter droplets. Scaffold-free cell spheroids and 3D tissue constructs recapitulate many aspects of functional human tissue more accurately than 2D or single cell cultures, but assembly methods for spheroids, e.g. hanging drop micro-plates, has had limited throughput. The increased throughput and decreased cost of our method enables spheroid production at the scale needed for lead discovery drug screening and approaches the cost where these micro tissues could be used as building blocks for organ scale regenerative medicine.

National Category
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy) Pharmaceutical Biotechnology Biomedical Laboratory Science/Technology
Identifiers
urn:nbn:se:kth:diva-250437 (URN)10.1101/552687 (DOI)
Note

QCR 20190617

Available from: 2019-04-29 Created: 2019-04-29 Last updated: 2019-06-17Bibliographically approved
Langer, K. & Jönsson, H. (2019). Rapid production and recovery of cell spheroids by automated droplet microfluidics. bioRxiv
Open this publication in new window or tab >>Rapid production and recovery of cell spheroids by automated droplet microfluidics
2019 (English)In: bioRxivArticle in journal (Refereed) Published
Abstract [en]

Droplet microfluidics enables high throughput cell processing, analysis and screening by miniaturizing the reaction vessels to nano- or pico-liter water-in oil droplets, but like many other microfluidic formats, droplet microfluidics have not been interfaced with or automated by laboratory robotics. Here we demonstrate automation of droplet microfluidics based on an inexpensive liquid handling robot for the automated production of human scaffold-free cell spheroids, using pipette actuation and interfacing the pipetting tip with a droplet generating microfluidic chip. In this chip we produce highly mono-disperse 290μm droplets with diameter CV of 1.7%. By encapsulating cells in these droplets, we produce cell spheroids in droplets and recover them to standard formats at a throughput of 85000 spheroids per microfluidic circuit per hour. The viability of the cells in spheroids remains high after recovery only decreased by 4% starting from 96% after 16 hours incubation in nanoliter droplets. Scaffold-free cell spheroids and 3D tissue constructs recapitulate many aspects of functional human tissue more accurately than 2D or single cell cultures, but assembly methods for spheroids, e.g. hanging drop micro-plates, has had limited throughput. The increased throughput and decreased cost of our method enables spheroid production at the scale needed for lead discovery drug screening and approaches the cost where these micro tissues could be used as building blocks for organ scale regenerative medicine.

Place, publisher, year, edition, pages
Cold Spring Harbor Laboratory, 2019
National Category
Pharmaceutical Biotechnology Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy) Medical Laboratory and Measurements Technologies
Identifiers
urn:nbn:se:kth:diva-250382 (URN)10.1101/552687 (DOI)
Note

QC 20190624

Available from: 2019-04-29 Created: 2019-04-29 Last updated: 2019-06-24Bibliographically approved
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
Huang, M., Jönsson, H. & Nielsen, J. (2018). High-throughput microfluidics for the screening of yeast libraries. In: Synthetic Metabolic Pathways: Methods and Protocols (pp. 307-317). Humana Press, 1671
Open this publication in new window or tab >>High-throughput microfluidics for the screening of yeast libraries
2018 (English)In: Synthetic Metabolic Pathways: Methods and Protocols, Humana Press, 2018, Vol. 1671, p. 307-317Chapter in book (Refereed)
Abstract [en]

Cell factory development is critically important for efficient biological production of chemicals, biofuels, and pharmaceuticals. Many rounds of the Design–Build–Test–Learn cycles may be required before an engineered strain meeting specific metrics required for industrial application. The bioindustry prefer products in secreted form (secreted products or extracellular metabolites) as it can lower the cost of downstream processing, reduce metabolic burden to cell hosts, and allow necessary modification on the final products, such as biopharmaceuticals. Yet, products in secreted form result in the disconnection of phenotype from genotype, which may have limited throughput in the Test step for identification of desired variants from large libraries of mutant strains. In droplet microfluidic screening, single cells are encapsulated in individual droplet and enable high-throughput processing and sorting of single cells or clones. Encapsulation in droplets allows this technology to overcome the throughput limitations present in traditional methods for screening by extracellular phenotypes. In this chapter, we describe a protocol/guideline for high-throughput droplet microfluidics screening of yeast libraries for higher protein secretion. This protocol can be adapted to screening by a range of other extracellular products from yeast or other hosts.

Place, publisher, year, edition, pages
Humana Press, 2018
Series
Methods in Molecular Biology, ISSN 1064-3745 ; 1671
Keywords
Droplet microfluidics, High-throughput screening, Protein secretion, Random mutagenesis, Systems biology, Yeast cell factories
National Category
Biological Sciences
Identifiers
urn:nbn:se:kth:diva-219635 (URN)10.1007/978-1-4939-7295-1_19 (DOI)29170967 (PubMedID)2-s2.0-85035111151 (Scopus ID)978-1-4939-7294-4 (ISBN)
Note

QC 20191025

Available from: 2017-12-11 Created: 2017-12-11 Last updated: 2019-10-25Bibliographically approved
Siedler, S., Khatri, N. K., Zsohár, A., Kjærbølling, I., Vogt, M., Hammar, P., . . . Jönsson, H. (2017). Development of a Bacterial Biosensor for Rapid Screening of Yeast p-Coumaric Acid Production. ACS Synthetic Biology, 6(10), 1860-1869
Open this publication in new window or tab >>Development of a Bacterial Biosensor for Rapid Screening of Yeast p-Coumaric Acid Production
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2017 (English)In: ACS Synthetic Biology, E-ISSN 2161-5063, Vol. 6, no 10, p. 1860-1869Article in journal (Refereed) Published
Abstract [en]

Transcription factor-based biosensors are used to identify producer strains, a critical bottleneck in cell factory engineering. Here, we address two challenges with this methodology: transplantation of heterologous transcriptional regulators into new hosts to generate functional biosensors and biosensing of the extracellular product concentration that accurately reflects the effective cell factory production capacity. We describe the effects of different translation initiation rates on the dynamic range of a p-coumaric acid biosensor based on the Bacillus subtilis transcriptional repressor PadR by varying its ribosomal binding site. Furthermore, we demonstrate the functionality of this p-coumaric acid biosensor in Escherichia coli and Corynebacterium glutamicum. Finally, we encapsulate yeast p-coumaric acid-producing cells with E. coli-biosensing cells in picoliter droplets and, in a microfluidic device, rapidly sort droplets containing yeast cells producing high amounts of extracellular p-coumaric acid using the fluorescent E. coli biosensor signal. As additional biosensors become available, such approaches will find broad applications for screening of an extracellular product.

Place, publisher, year, edition, pages
American Chemical Society (ACS), 2017
Keywords
Para coumaric acid, recombinant DNA, bacterial protein, propionic acid derivative, trans-3-(4'-hydroxyphenyl)-2-propenoic acid, transcription factor, article, bacillus subtilis, bacterium culture, binding site, catalysis, cell population, cell selection, coculture, controlled study, Corynebacterium glutamicum, Escherichia coli, feedback system, fluorescence, fungal cell culture, gene repression, Lactococcus lactis, microbiological examination, microencapsulation, microfluidics, mutagenesis, nonhuman, osmosis, phenotype, priority journal, promoter region, translation initiation, water transport, yeast, yeast cell, genetic procedures, metabolism, procedures, Bacterial Proteins, Biosensing Techniques, Propionates, Transcription Factors
National Category
Cell Biology Other Industrial Biotechnology Biocatalysis and Enzyme Technology
Identifiers
urn:nbn:se:kth:diva-227082 (URN)10.1021/acssynbio.7b00009 (DOI)000413715200008 ()28532147 (PubMedID)2-s2.0-85031721311 (Scopus ID)
Note

QC 20191029

Available from: 2018-05-16 Created: 2018-05-16 Last updated: 2019-10-29Bibliographically approved
Siedler, S., Khatri, N. K., Zsohár, A., Kjærbølling, I., Vogt, M., Hammar, P., . . . Jönsson, H. (2016). High throughput droplet sorting of yeast for p-Coumaric acid production detected by co-encapsulated E. coli biosensor bacteria. In: 20th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2016: . Paper presented at 20th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2016, 9 October 2016 through 13 October 2016 (pp. 551-552). Chemical and Biological Microsystems Society
Open this publication in new window or tab >>High throughput droplet sorting of yeast for p-Coumaric acid production detected by co-encapsulated E. coli biosensor bacteria
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2016 (English)In: 20th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2016, Chemical and Biological Microsystems Society , 2016, p. 551-552Conference paper, Published paper (Refereed)
Abstract [en]

We present the optimization of E. coli biosensor bacteria and demonstrate their use to detect and sort co-encapsulated S. cerevisiae yeast cells by their p-Coumaric acid (pCA) production in picoliter microfluidic droplets at high throughput. This strategy allows us to enrich pCA producing cell factories by the cell permeable product pCA using a biosensor in a separate cell.

Place, publisher, year, edition, pages
Chemical and Biological Microsystems Society, 2016
Keywords
Biosensors, Droplet microfluidics, High throughput screening, Synthetic biology
National Category
Biological Sciences
Identifiers
urn:nbn:se:kth:diva-207552 (URN)2-s2.0-85014274649 (Scopus ID)9780979806490 (ISBN)
Conference
20th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2016, 9 October 2016 through 13 October 2016
Note

Conference code: 126047; Export Date: 22 May 2017; Conference Paper; Correspondence Address: Joensson, H.N.; KTH Royal Institute of TechnologySweden; email: hakan.jonsson@biotech.kth.se. QC 20170531

Available from: 2017-05-31 Created: 2017-05-31 Last updated: 2017-05-31Bibliographically approved
Afrasiabi, R., Söderberg, L. M., Jönsson, H., Björk, P., Svahn Andersson, H. & Linnros, J. (2016). Integration of a Droplet-Based Microfluidic System and Silicon Nanoribbon FET Sensor. Micromachines, 7(8)
Open this publication in new window or tab >>Integration of a Droplet-Based Microfluidic System and Silicon Nanoribbon FET Sensor
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2016 (English)In: Micromachines, ISSN 2072-666X, E-ISSN 2072-666X, Vol. 7, no 8Article in journal (Refereed) Published
Abstract [en]

We present a novel microfluidic system that integrates droplet microfluidics with a silicon nanoribbon field-effect transistor (SiNR FET), and utilize this integrated system to sense differences in pH. The device allows for selective droplet transfer to a continuous water phase, actuated by dielectrophoresis, and subsequent detection of the pH level in the retrieved droplets by SiNR FETs on an electrical sensor chip. The integrated microfluidic system demonstrates a label-free detection method for droplet microfluidics, presenting an alternative to optical fluorescence detection. In this work, we were able to differentiate between droplet trains of one pH-unit difference. The pH-based detection method in our integrated system has the potential to be utilized in the detection of biochemical reactions that induce a pH-shift in the droplets.

Place, publisher, year, edition, pages
MDPI AG, 2016
Keywords
NanoFET; silicon nanoribbon; droplet microfluidics; pH measurement
National Category
Nano Technology
Research subject
Information and Communication Technology
Identifiers
urn:nbn:se:kth:diva-191177 (URN)10.3390/mi7080134 (DOI)000382467700006 ()2-s2.0-84984791952 (Scopus ID)
Funder
Knut and Alice Wallenberg Foundation
Note

QC 20160825. QC 20191126

Available from: 2016-08-25 Created: 2016-08-25 Last updated: 2019-11-26Bibliographically approved
Fornell, A., Nilsson, J., Jonsson, L., Periyannan Rajeswari, P. K., Jönsson, H. & Tenje, M. (2016). Particle enrichment in droplet acoustofluidics. In: Micronano System Workshop (MSW 2016), Lund, Sweden, May 17-18 2016: . Paper presented at Micronano System Workshop (MSW 2016), Lund, Sweden, May 17-18 2016.
Open this publication in new window or tab >>Particle enrichment in droplet acoustofluidics
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2016 (English)In: Micronano System Workshop (MSW 2016), Lund, Sweden, May 17-18 2016, 2016Conference paper, Oral presentation with published abstract (Refereed)
National Category
Analytical Chemistry Medical Laboratory and Measurements Technologies
Identifiers
urn:nbn:se:kth:diva-250372 (URN)
Conference
Micronano System Workshop (MSW 2016), Lund, Sweden, May 17-18 2016
Note

QCR 20190624

Available from: 2019-04-29 Created: 2019-04-29 Last updated: 2019-06-24Bibliographically approved
Fornell, A., Nilsson, J., Jonsson, L., Periyannan Rajeswari, P. K., Jönsson, H. & Tenje, M. (2016). Particle enrichment in two-phase microfluidic systems using acoustophoresis. In: Acoustofluidics 2016, Kongens Lyngby, Denmark, September 22-23 2016: . Paper presented at 20th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2016.
Open this publication in new window or tab >>Particle enrichment in two-phase microfluidic systems using acoustophoresis
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2016 (English)In: Acoustofluidics 2016, Kongens Lyngby, Denmark, September 22-23 2016, 2016Conference paper, Poster (with or without abstract) (Refereed)
National Category
Other Electrical Engineering, Electronic Engineering, Information Engineering Biomedical Laboratory Science/Technology Analytical Chemistry
Identifiers
urn:nbn:se:kth:diva-250374 (URN)
Conference
20th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2016
Note

QC 20190819

Available from: 2019-04-29 Created: 2019-04-29 Last updated: 2019-08-19Bibliographically approved
Organisations
Identifiers
ORCID iD: ORCID iD iconorcid.org/0000-0001-5232-0805

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