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A sensitive and real-time assay of trypsin by using molecular imprinting-based capacitive biosensor

Ertürk, Gizem LU ; Hedström, Martin LU and Mattiasson, Bo LU (2016) In Biosensors and Bioelectronics 86. p.557-565
Abstract

Use of a highly sensitive, selective capacitive biosensor is reported for label-free, real-time, easy and rapid detection of trypsin by using the microcontact imprinting method. Real-time trypsin detection was performed with trypsin-imprinted (trypsin-MIP) capacitive electrodes using standard trypsin solutions in the concentration range of 1.0×10−13–1.0×10−7 M with a detection limit of 3.0×10−13 M. Selectivity and cross-reactivity of the system were tested by using competing proteins including chymotrypsin (chy), bovine serum albumin (BSA), lysozyme (lyz) and cytochrome c (cyt c) in singular and competitive manner and the selectivity of the system was determined with the selectivity coefficients of... (More)

Use of a highly sensitive, selective capacitive biosensor is reported for label-free, real-time, easy and rapid detection of trypsin by using the microcontact imprinting method. Real-time trypsin detection was performed with trypsin-imprinted (trypsin-MIP) capacitive electrodes using standard trypsin solutions in the concentration range of 1.0×10−13–1.0×10−7 M with a detection limit of 3.0×10−13 M. Selectivity and cross-reactivity of the system were tested by using competing proteins including chymotrypsin (chy), bovine serum albumin (BSA), lysozyme (lyz) and cytochrome c (cyt c) in singular and competitive manner and the selectivity of the system was determined with the selectivity coefficients of approximately 705.1, 6.5, 6.4 and 5.1 for chy, BSA, lyz and cyt c, respectively. The trypsin-MIP capacitive electrode was used for ~80 assays during 2 months and retained its binding property during all that time with a decrease of approximately 2.3% in the signal amplitude. In the last step, trypsin activity was measured by using Nα-Benzoyl-D, L-arginine 4-nitroanilide hydrochloride (BAPNA) as the substrate with spectrophotometer at 410 nm. The trypsin activity was measured as 9 mU/mL by spectrophotometer while the amount of captured enzyme calculated from the capacitive system was 7.9 mU/mL which shows the correlation between two methods. From the comparison it is obvious that the new method is an attractive alternative for assaying trypsin and the developed capacitive system might be used successfully to monitor label-free, real-time enzymatic activity of different proteases in a sensitive, rapid, cost-effective manner for different applications.

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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Capacitive biosensor, Cross-reactivity, Microcontact imprinting, Selectivity
in
Biosensors and Bioelectronics
volume
86
pages
9 pages
publisher
Elsevier
external identifiers
  • Scopus:84989892993
ISSN
0956-5663
DOI
10.1016/j.bios.2016.07.046
language
English
LU publication?
yes
id
ea6d043a-01e3-4a01-a28e-10cdf047e6f2
date added to LUP
2016-10-18 08:10:02
date last changed
2016-10-19 03:00:05
@misc{ea6d043a-01e3-4a01-a28e-10cdf047e6f2,
  abstract     = {<p>Use of a highly sensitive, selective capacitive biosensor is reported for label-free, real-time, easy and rapid detection of trypsin by using the microcontact imprinting method. Real-time trypsin detection was performed with trypsin-imprinted (trypsin-MIP) capacitive electrodes using standard trypsin solutions in the concentration range of 1.0×10<sup>−13</sup>–1.0×10<sup>−7</sup> M with a detection limit of 3.0×10<sup>−13</sup> M. Selectivity and cross-reactivity of the system were tested by using competing proteins including chymotrypsin (chy), bovine serum albumin (BSA), lysozyme (lyz) and cytochrome c (cyt c) in singular and competitive manner and the selectivity of the system was determined with the selectivity coefficients of approximately 705.1, 6.5, 6.4 and 5.1 for chy, BSA, lyz and cyt c, respectively. The trypsin-MIP capacitive electrode was used for ~80 assays during 2 months and retained its binding property during all that time with a decrease of approximately 2.3% in the signal amplitude. In the last step, trypsin activity was measured by using N<sub>α</sub>-Benzoyl-D, L-arginine 4-nitroanilide hydrochloride (BAPNA) as the substrate with spectrophotometer at 410 nm. The trypsin activity was measured as 9 mU/mL by spectrophotometer while the amount of captured enzyme calculated from the capacitive system was 7.9 mU/mL which shows the correlation between two methods. From the comparison it is obvious that the new method is an attractive alternative for assaying trypsin and the developed capacitive system might be used successfully to monitor label-free, real-time enzymatic activity of different proteases in a sensitive, rapid, cost-effective manner for different applications.</p>},
  author       = {Ertürk, Gizem and Hedström, Martin and Mattiasson, Bo},
  issn         = {0956-5663},
  keyword      = {Capacitive biosensor,Cross-reactivity,Microcontact imprinting,Selectivity},
  language     = {eng},
  month        = {12},
  pages        = {557--565},
  publisher    = {ARRAY(0x6cc7bc0)},
  series       = {Biosensors and Bioelectronics},
  title        = {A sensitive and real-time assay of trypsin by using molecular imprinting-based capacitive biosensor},
  url          = {http://dx.doi.org/10.1016/j.bios.2016.07.046},
  volume       = {86},
  year         = {2016},
}