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

Research output: Contribution to journalArticle

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 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.

Details

Authors
Organisations
External organisations
  • CapSenze HB
  • CapSenze Biosystems AB
Research areas and keywords

Subject classification (UKÄ) – MANDATORY

  • Analytical Chemistry

Keywords

  • Capacitive biosensor, Cross-reactivity, Microcontact imprinting, Selectivity
Original languageEnglish
Pages (from-to)557-565
Number of pages9
JournalBiosensors and Bioelectronics
Volume86
Publication statusPublished - 2016 Dec 15
Publication categoryResearch
Peer-reviewedYes