eprintid: 1472255
rev_number: 22
eprint_status: archive
userid: 608
dir: disk0/01/47/22/55
datestamp: 2015-11-06 11:35:20
lastmod: 2021-10-04 01:45:48
status_changed: 2015-11-06 11:35:20
type: article
metadata_visibility: show
creators_name: Kukwikila, M
creators_name: Howorka, S
title: Nanopore-Based Electrical and Label-Free Sensing of Enzyme Activity in Blood Serum
ispublished: pub
divisions: UCL
divisions: B04
divisions: C06
divisions: F56
note: © 2015 American Chemical Society. This is an open access article published under a Creative Commons Attribution (CC-BY)
License, which permits unrestricted use, distribution and reproduction in any medium, provided the author and source are cited.
abstract: A generic strategy to expand the analytical scope of electrical nanopore sensing is presented. We specifically and electrically detect the activity of a diagnostically relevant hydrolytic enzyme and remove the analytically harmful interference from the biochemically complex sample matrix of blood serum. Our strategy is demonstrated at the example of the renin protease which is involved in regulation of blood pressure. The analysis scheme exploits a new approach to reduce sample complexity while generating a specific read-out signal. Within a single spin-column (i), the protease cleaves a resin-tethered peptide substrate (ii) which is affinity-purified using the same multifunctional resin to remove interfering blood serum components, followed by (iii) detecting the peptide via electrical nanopore recordings. Our approach is beneficial in several ways. First, by eliminating serum components, we overcome limitations of nanopore sensing when challenging samples lead to membrane instability and a poor signal-to-noise ratio. Second, the label-free sensing avoids drawbacks of currently used radiolabel-immunoassays for renin. Finally, the strategy of simultaneous generation and purification of a signal peptide within a multifunctional resin can very likely be expanded to other hydrolytic enzymes dissolved in any analyte matrix and exploited for analytical read-out methods other than nanopore sensing.
date: 2015-09-15
date_type: published
official_url: http://dx.doi.org/10.1021/acs.analchem.5b01764
oa_status: green
full_text_type: pub
language: eng
primo: open
primo_central: open_green
article_type_text: Journal Article
verified: verified_manual
elements_id: 1051059
doi: 10.1021/acs.analchem.5b01764
language_elements: eng
lyricists_name: Howorka, Stefan
lyricists_id: SHOWO39
actors_name: Howorka, Stefan
actors_name: Poirier, Elizabeth
actors_id: SHOWO39
actors_id: EPPOI23
actors_role: owner
actors_role: impersonator
full_text_status: public
publication: Anal Chem
volume: 87
number: 18
pagerange: 9149-9154
event_location: United States
issn: 1520-6882
citation:        Kukwikila, M;    Howorka, S;      (2015)    Nanopore-Based Electrical and Label-Free Sensing of Enzyme Activity in Blood Serum.                   Anal Chem , 87  (18)   pp. 9149-9154.    10.1021/acs.analchem.5b01764 <https://doi.org/10.1021/acs.analchem.5b01764>.       Green open access   
 
document_url: https://discovery.ucl.ac.uk/id/eprint/1472255/1/acs.analchem.5b01764.pdf