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Isothermal microcalorimetry accurately detects bacteria, tumorous microtissues, and parasitic worms in a label-free well-plate assay

Braissant, Olivier; Keiser, Jennifer; Meister, Isabel; Bachmann, Alexander; Wirz, Dieter; Goepfert, Beat; Bonkat, Gernot and Wadsö, Ingemar LU (2015) In Biotechnology Journal 10(3). p.460-468
Abstract
Isothermal microcalorimetry is a label-free assay that allows monitoring of enzymatic and metabolic activities. The technique has strengths, but most instruments have a low throughput, which has limited their use for bioassays. Here, an isothermal microcalorimeter, equipped with a vessel holder similar to a 48-well plate, was used. The increased throughput of this microcalorimeter makes it valuable for biomedical and pharmaceutical applications. Our results show that the sensitivity of the instrument allows the detection of 3 x 10(4) bacteria per vial. Growth of P. mirabilis in Luria Broth medium was detected between 2 and 9 h with decreasing inoculum. The culture released 2.1J with a maximum thermal power of 76 W. The growth rate... (More)
Isothermal microcalorimetry is a label-free assay that allows monitoring of enzymatic and metabolic activities. The technique has strengths, but most instruments have a low throughput, which has limited their use for bioassays. Here, an isothermal microcalorimeter, equipped with a vessel holder similar to a 48-well plate, was used. The increased throughput of this microcalorimeter makes it valuable for biomedical and pharmaceutical applications. Our results show that the sensitivity of the instrument allows the detection of 3 x 10(4) bacteria per vial. Growth of P. mirabilis in Luria Broth medium was detected between 2 and 9 h with decreasing inoculum. The culture released 2.1J with a maximum thermal power of 76 W. The growth rate calculated using calorimetric and spectrophotometric data were 0.60 and 0.57 h(-1), respectively. Additional insight on protease activities of P. mirabilis matching the last peak in heat production could be gathered as well. Growth of tumor microtissues releasing a maximum thermal power of 2.1 W was also monitored and corresponds to a diameter increase of the microtissues from ca. 100 to 428 m. This opens new research avenues in cancer research, diagnostics, and development of new antitumor drugs. For parasitic worms, the technique allows assessment of parasite survival using motor and metabolic activities even with a single worm. (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Analytical biotechnology, Cell biology, Diseases, Isothermal, microcalorimetry, Metabolic flux analysis
in
Biotechnology Journal
volume
10
issue
3
pages
460 - 468
publisher
John Wiley & Sons
external identifiers
  • wos:000351366600012
  • scopus:84924606514
ISSN
1860-6768
DOI
10.1002/biot.201400494
language
English
LU publication?
yes
id
e878eb9b-cbca-44dd-8774-28852df16e20 (old id 5293696)
date added to LUP
2015-04-24 14:53:22
date last changed
2017-09-17 03:48:49
@article{e878eb9b-cbca-44dd-8774-28852df16e20,
  abstract     = {Isothermal microcalorimetry is a label-free assay that allows monitoring of enzymatic and metabolic activities. The technique has strengths, but most instruments have a low throughput, which has limited their use for bioassays. Here, an isothermal microcalorimeter, equipped with a vessel holder similar to a 48-well plate, was used. The increased throughput of this microcalorimeter makes it valuable for biomedical and pharmaceutical applications. Our results show that the sensitivity of the instrument allows the detection of 3 x 10(4) bacteria per vial. Growth of P. mirabilis in Luria Broth medium was detected between 2 and 9 h with decreasing inoculum. The culture released 2.1J with a maximum thermal power of 76 W. The growth rate calculated using calorimetric and spectrophotometric data were 0.60 and 0.57 h(-1), respectively. Additional insight on protease activities of P. mirabilis matching the last peak in heat production could be gathered as well. Growth of tumor microtissues releasing a maximum thermal power of 2.1 W was also monitored and corresponds to a diameter increase of the microtissues from ca. 100 to 428 m. This opens new research avenues in cancer research, diagnostics, and development of new antitumor drugs. For parasitic worms, the technique allows assessment of parasite survival using motor and metabolic activities even with a single worm.},
  author       = {Braissant, Olivier and Keiser, Jennifer and Meister, Isabel and Bachmann, Alexander and Wirz, Dieter and Goepfert, Beat and Bonkat, Gernot and Wadsö, Ingemar},
  issn         = {1860-6768},
  keyword      = {Analytical biotechnology,Cell biology,Diseases,Isothermal,microcalorimetry,Metabolic flux analysis},
  language     = {eng},
  number       = {3},
  pages        = {460--468},
  publisher    = {John Wiley & Sons},
  series       = {Biotechnology Journal},
  title        = {Isothermal microcalorimetry accurately detects bacteria, tumorous microtissues, and parasitic worms in a label-free well-plate assay},
  url          = {http://dx.doi.org/10.1002/biot.201400494},
  volume       = {10},
  year         = {2015},
}