Sorting catalytically active polymersome nanoreactors by flow cytometry
Flow cytometry is a powerful technique for high-throughput,...
| Main Authors: | , , , , , , , |
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| Other Authors: | |
| Format: | Journal Article |
| Language: | English |
| Published: |
2011
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| Subjects: | |
| Online Access: | https://hdl.handle.net/10356/93845 http://hdl.handle.net/10220/7282 |
| _version_ | 1811681940639580160 |
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| author | Nallani, Madhavan Woestenenk, Rob Hans-Peter M. de Hoog Dongen, Stijn F. M. van Boezeman, Jan Cornelissen, Jeroen J. L. M. Nolte, Roeland J. M. Hest, Jan C. M. van. |
| author2 | School of Materials Science & Engineering |
| author_facet | School of Materials Science & Engineering Nallani, Madhavan Woestenenk, Rob Hans-Peter M. de Hoog Dongen, Stijn F. M. van Boezeman, Jan Cornelissen, Jeroen J. L. M. Nolte, Roeland J. M. Hest, Jan C. M. van. |
| author_sort | Nallani, Madhavan |
| collection | NTU |
| description | Flow cytometry is a powerful technique for high-throughput,
fluorescence-activated screening and sorting of cells (FACS).
This methodology has been extended by Griffiths to the
screening of water-in-oil microdroplets filled with an in vitro
protein expression system.[1–6] The catalytic gene product was
detected by the transformation of a co-encapsulated profluorescent
substrate into a fluorescent product. Here we
report a strategy that involves a versatile one-step preparation
procedure of enzyme filled porous and stable polymeric
capsules (polymersomes). Since the pores of the capsules are
small enough to keep enzymes in, whereas these are
sufficiently large to allow (profluorescent) substrates to enter,
enzyme activity screening can be performed by the build-up of
fluorescence, followed by FACS. To prevent the substrate
from diffusing out of the capsules, a trapping agent was added
inside the capsule. With this technology we were able to
separate enzymatically active polymersomes from non-filled
or non-active polymersomes. |
| first_indexed | 2024-10-01T03:48:56Z |
| format | Journal Article |
| id | ntu-10356/93845 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T03:48:56Z |
| publishDate | 2011 |
| record_format | dspace |
| spelling | ntu-10356/938452020-06-01T10:26:30Z Sorting catalytically active polymersome nanoreactors by flow cytometry Nallani, Madhavan Woestenenk, Rob Hans-Peter M. de Hoog Dongen, Stijn F. M. van Boezeman, Jan Cornelissen, Jeroen J. L. M. Nolte, Roeland J. M. Hest, Jan C. M. van. School of Materials Science & Engineering DRNTU::Engineering::Materials::Biomaterials Flow cytometry is a powerful technique for high-throughput, fluorescence-activated screening and sorting of cells (FACS). This methodology has been extended by Griffiths to the screening of water-in-oil microdroplets filled with an in vitro protein expression system.[1–6] The catalytic gene product was detected by the transformation of a co-encapsulated profluorescent substrate into a fluorescent product. Here we report a strategy that involves a versatile one-step preparation procedure of enzyme filled porous and stable polymeric capsules (polymersomes). Since the pores of the capsules are small enough to keep enzymes in, whereas these are sufficiently large to allow (profluorescent) substrates to enter, enzyme activity screening can be performed by the build-up of fluorescence, followed by FACS. To prevent the substrate from diffusing out of the capsules, a trapping agent was added inside the capsule. With this technology we were able to separate enzymatically active polymersomes from non-filled or non-active polymersomes. 2011-10-17T03:26:29Z 2019-12-06T18:46:32Z 2011-10-17T03:26:29Z 2019-12-06T18:46:32Z 2008 2008 Journal Article Nallani, M., Woestenenk, R., Hoog, H. P. M. D., Dongen, S. F. M. V., Boezeman, J., Cornelissen, J. J. L. M., et al. (2009). Sorting Catalytically Active Polymersome Nanoreactors by Flow Cytometry. Small, 5(10), 1138-1143. 1613-6810 https://hdl.handle.net/10356/93845 http://hdl.handle.net/10220/7282 10.1002/smll.200801204 155293 en Small © 2009 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim. 6 p. |
| spellingShingle | DRNTU::Engineering::Materials::Biomaterials Nallani, Madhavan Woestenenk, Rob Hans-Peter M. de Hoog Dongen, Stijn F. M. van Boezeman, Jan Cornelissen, Jeroen J. L. M. Nolte, Roeland J. M. Hest, Jan C. M. van. Sorting catalytically active polymersome nanoreactors by flow cytometry |
| title | Sorting catalytically active polymersome nanoreactors by flow cytometry |
| title_full | Sorting catalytically active polymersome nanoreactors by flow cytometry |
| title_fullStr | Sorting catalytically active polymersome nanoreactors by flow cytometry |
| title_full_unstemmed | Sorting catalytically active polymersome nanoreactors by flow cytometry |
| title_short | Sorting catalytically active polymersome nanoreactors by flow cytometry |
| title_sort | sorting catalytically active polymersome nanoreactors by flow cytometry |
| topic | DRNTU::Engineering::Materials::Biomaterials |
| url | https://hdl.handle.net/10356/93845 http://hdl.handle.net/10220/7282 |
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