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Ultrafast molecular motor driven nanoseparation and biosensing
The Nanometer Structure Consortium (nmC@LU), Division of Solid State Physics, Lund University. (Heiner Linke)
Linnaeus University, Faculty of Health and Life Sciences, Department of Chemistry and Biomedical Sciences. (Alf Månsson)ORCID iD: 0000-0001-6878-3142
Linnaeus University, Faculty of Health and Life Sciences, Department of Chemistry and Biomedical Sciences. (Alf Månsson)
Linnaeus University, Faculty of Health and Life Sciences, Department of Chemistry and Biomedical Sciences. (Alf Månsson)ORCID iD: 0000-0003-2819-3046
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2013 (English)In: Biosensors & bioelectronics, ISSN 0956-5663, E-ISSN 1873-4235, Vol. 48, 145-152 p.Article in journal (Refereed) Published
Abstract [en]

Portable biosensor systems would benefit from reduced dependency on external power supplies as well as from further miniaturization and increased detection rate. Systems built around self-propelled biological molecular motors and cytoskeletal filaments hold significant promise in these regards as they are built from nanoscale components that enable nanoseparation independent of fluidic pumping. Previously reported microtubule-kinesin based devices are slow, however, compared to several existing biosensor systems. Here we demonstrate that this speed limitation can be overcome by using the faster actomyosin motor system. Moreover, due to lower flexural rigidity of the actin filaments, smaller features can be achieved compared to microtubule-based systems, enabling further miniaturization. Using a device designed through optimization by Monte Carlo simulations, we demonstrate extensive myosin driven enrichment of actin filaments on a detector area of less than 10 μm2, with a concentration half-time of approximately 40 s. We also show accumulation of model analyte (streptavidin at nanomolar concentration in nanoliter effective volume) detecting increased fluorescence intensity within seconds after initiation of motor-driven transportation from capture regions. We discuss further optimizations of the system and incorporation into a complete biosensing workflow.

Place, publisher, year, edition, pages
Elsevier, 2013. Vol. 48, 145-152 p.
Keyword [en]
Actin filament; Diagnostics; Electron beam lithography; Heavy meromyosin; Nanoseparation; Monte-Carlo simulation
National Category
Biochemistry and Molecular Biology
Research subject
Natural Science, Biomedical Sciences
Identifiers
URN: urn:nbn:se:lnu:diva-28382DOI: 10.1016/j.bios.2013.03.071ISI: 000321085600024Scopus ID: 2-s2.0-84877888579OAI: oai:DiVA.org:lnu-28382DiVA: diva2:642575
Funder
EU, FP7, Seventh Framework Programme, 228971The nanometer Structure Consortium at Lund UniversitySwedish Research CouncilKnut and Alice Wallenberg Foundation
Available from: 2013-08-22 Created: 2013-08-22 Last updated: 2017-12-06Bibliographically approved
In thesis
1. Towards Myosin Powered Lab-on-a-Chip Devices
Open this publication in new window or tab >>Towards Myosin Powered Lab-on-a-Chip Devices
2013 (English)Doctoral thesis, comprehensive summary (Other academic)
Alternative title[sv]
Mot utvecklandet av myosindrivna laboratorier på chip
Abstract [en]

Myosins are protein motors that use chemical energy in the form of adenosinetriphosphate to produce force and motion. These molecular motors might be usedto power transportation in Lab-on-a-chip devices where a series of laboratory tasks(e.g. separation, concentration and detection) are performed in one sequence on asmall chip. Because of the small size, lab-on-a-chip devices are predicted to befaster and more sensitive than conventional systems. Further potential advantagesinclude cost efficiency and the possibility to perform many analyzes in parallel.Substituting microfluidics with myosin based transport would allow furtherminiaturization and make lab-on-a-chip devices more readily portable by reducingthe need for external power supplies. However, there are also limitations thathamper the development of such devices. Here we investigate several aspects of amyosin powered lab-on-a-chip device and present ways to overcome criticallimitations. First we demonstrate covalent attachment of antibodies to actinfilament shuttles with retained ability of the filaments to be propelled by myosinfragments, previously believed to be difficult. Secondly we develop a separationmethod to overcome the deleterious effects of body fluids on the actomyosinsystem. Thirdly, we explore the possibility to concentrate actin shuttles on ananostructured surface and achieve >20 times concentration in <1 min. Monte-Carlo simulations of the concentration process suggest further room forimprovement. Fourth, we develop novel techniques for fast and automaticdetection of fluorescence at certain check points which improves S/N ratio >20times. Finally, we take the first steps towards the development of threedimensional,nanowire-based transport systems, important both for lab-on-a-chipapplications and fundamental studies. Our results demonstrate the potential of amyosin based lab-on-a-chip device and lay the foundation for furtherdevelopments. Thus, we anticipate that this work will influence future studiestowards a complete diagnostic lab-on-a-chip work-up based on molecular motors.In addition, the work might also have implications for the development of futurebiocomputation and drug screening devices as well as novel biophysical studies ofthe actomyosin system.

Place, publisher, year, edition, pages
Växjö: Linnaeus University Press, 2013. 200 p.
Series
Linnaeus University Dissertations, 144/2013
Keyword
myosin, actin, molecular motors, lab-on-a-chip, nanobiotechnology, bionanotechnology, diagnostics, nanowires, nanowire arrays
National Category
Biochemistry and Molecular Biology
Research subject
Natural Science, Biomedical Sciences
Identifiers
urn:nbn:se:lnu:diva-28384 (URN)978-91-87427-45-9 (ISBN)
Public defence
2013-09-13, N2007, Västergård, Smålandsgatan 26E, Kalmar, 09:00 (English)
Opponent
Supervisors
Funder
EU, FP7, Seventh Framework Programme, 228971
Available from: 2013-09-10 Created: 2013-08-22 Last updated: 2016-05-03Bibliographically approved

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ten Siethoff, LasseKumar, SarojPersson, MalinMånsson, Alf

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