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Measurement of prot...
Measurement of protein binding with vastly improved time resolution using a quartz crystal microbalance driven at a fixed frequency
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- Guha, Arnab (författare)
- Wolfson School of Mechanical, Electrical and Manufacturing Engineering, Epinal Way, Loughborough, Loughborough University, LE11 3TU, UK
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- Sandström, Niklas, 1981- (författare)
- KTH,Biofysik,Önfelt Lab
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- Ostanin, Victor (författare)
- Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge, CB2 1EW , UK
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visa fler...
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- van der Wijngaart, Wouter (författare)
- KTH,Mikro- och nanosystemteknik
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- Klenerman, David (författare)
- Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge, CB2 1EW , UK
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- Ghosh, Sourav (författare)
- Wolfson School of Mechanical, Electrical and Manufacturing Engineering, Epinal Way, Loughborough, Loughborough University, LE11 3TU, UK
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visa färre...
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(creator_code:org_t)
- 2017
- 2017
- Engelska.
- Relaterad länk:
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https://urn.kb.se/re...
Abstract
Ämnesord
Stäng
- Introduction: Quartz crystal microbalance (QCM) is commonly used to study biomolecular binding by measuring shifts in resonance frequency of a quartz-crystal-oscillator. However, the currently used methods like impedance analysis or QCM-D, which require repeated sweeps or ringing, are limited in time resolution (~1 second) due to the need for averaging. This restricts our ability to study transient biomolecular processes, which occur in sub-millisecond time scale. A novel technique has been reported here that allows quantification of resonance frequency of a quartz-crystal-oscillator with significantly improved time resolution by driving and measuring continuously at a constant frequency within the resonance bandwidth. Method: The reactive component of the experimentally obtained impedance is utilized for the estimation of resonance frequency from the Butterworth Van-dyke (BVD) model of a quartz-crystal-oscillator, assuming that changes in motional inductance and capacitance around resonance are negligible. Triplicate sets of experiments involving the binding of streptavidin with a biotin functionalized 14.3 MHz quartz oscillator surface were performed. Intermittent frequency sweeps and fixed frequency drives, both of 0.1 second duration and around 14.3 MHz, were taken at intervals of 2 minutes under the flow of phosphate-buffer-saline (PBS buffer) before and after injection of streptavidin. Results: The average shift in resonance frequency from the baseline (measurements before streptavidin injection) due to streptavidin-biotin binding, calculated from the fixed frequency drive or FFD (148 Hz) was within 1% of that estimated from the frequency sweep method by fitting the experimentally recorded impedance employing the BVD model (149 Hz). Discussion: The agreement of the FFD with conventional frequency sweep method suggests that protein binding can be quantified with reasonable accuracy from each impedance data point, which with our set-up is recorded at 30 kHz sampling rate. This gives a time resolution of 0.03 millisecond, which is about 4 orders of magnitude improvement over the state-of-the-art.
Ämnesord
- TEKNIK OCH TEKNOLOGIER -- Medicinteknik (hsv//swe)
- ENGINEERING AND TECHNOLOGY -- Medical Engineering (hsv//eng)
Nyckelord
- Bioteknologi
- Biotechnology
Publikations- och innehållstyp
- vet (ämneskategori)
- kon (ämneskategori)