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Rapid signal enhancement method for nanoprobe-based biosensing
KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology.
KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology.
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2017 (English)In: Scientific Reports, ISSN 2045-2322, E-ISSN 2045-2322, Vol. 7, article id 6837Article in journal (Refereed) Published
Abstract [en]

The introduction of nanomaterials as detection reagents has enabled improved sensitivity and facilitated detection in a variety of bioanalytical assays. However, high nanoprobe densities are typically needed for colorimetric detection and to circumvent this limitation several enhancement protocols have been reported. Nevertheless, there is currently a lack of universal, enzyme-free and versatile methods that can be readily applied to existing as well as new biosensing strategies. The novel method presented here is shown to enhance the signal of gold nanoparticles enabling visual detection of a spot containing < 10 nanoparticles. Detection of Protein G on paper arrays was improved by a 100-fold amplification factor in under five minutes of assay time, using IgG-labelled gold, silver, silica and iron oxide nanoprobes. Furthermore, we show that the presented protocol can be applied to a commercial allergen microarray assay, ImmunoCAP ISAC sIgE 112, attaining a good agreement with fluorescent detection when analysing human clinical samples.

Place, publisher, year, edition, pages
NATURE PUBLISHING GROUP , 2017. Vol. 7, article id 6837
National Category
Analytical Chemistry
Identifiers
URN: urn:nbn:se:kth:diva-212341DOI: 10.1038/s41598-017-07030-0ISI: 000406610000088Scopus ID: 2-s2.0-85026428495OAI: oai:DiVA.org:kth-212341DiVA, id: diva2:1135494
Note

QC 20170823

Available from: 2017-08-23 Created: 2017-08-23 Last updated: 2019-02-20Bibliographically approved
In thesis
1. Novel planar and particle-based microarrays for point-of-care diagnostics
Open this publication in new window or tab >>Novel planar and particle-based microarrays for point-of-care diagnostics
2019 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Point-of-care assays are easy-to-use, portable and inexpensive tests that can

be used to aid diagnostics by measuring levels of disease-specific molecules

in settings where access to advanced laboratory equipment and trained

personnel are limited, such as at the patient's bedside or in low resource

parts of developing countries. In order to achieve high multiplexing

capacities, such assays can be based on planar microarrays consisting of

spots immobilized on a flat surface or on particle-based microarrays based

on populations of encoded particles. The aim of the work presented in this

thesis is to develop new point-of-care amenable planar and particle-based

microarrays that allow for highly multiplexed assays while maintaining low

sample-to-result times, complexity and instrumentation requirements.

Paper I demonstrates the use graphically encoded particles for colorimetric

detection of autoantibodies using a consumer-grade flatbed scanner. Four

graphical characters on the surface of each particle allows for millions of

codes and the use of gold nanoparticles as a detection label allows both the

code and the signal intensity to be read out in a single image.

Paper II describes a signal enhancement method that increases the

sensitivity of gold nanoparticle detection on planar microarrays. Using this

method, detection of allergen-specific IgE can be carried out using a

consumer-grade flatbed scanner instead of a more expensive fluorescence

scanner without sacrificing assay performance.

Paper III demonstrates the use of an isothermal DNA amplification method

for detection of adenoviral DNA on a paper-based microarray. Using an

isothermal amplification method eliminates the need for a thermocycler,

reducing the instrumentation required for such detection.

Paper IV shows the use of solid-phase PCR to amplify bacterial DNA directly

on the surface of particles. This strategy reduces assay time by eliminating

the need for separate amplification and hybridisation steps.

Place, publisher, year, edition, pages
Stockholm: KTH Royal Institute of Technology, 2019. p. 88
Series
TRITA-CBH-FOU ; 2019:13
Keywords
Planar microarrays, Particle-based microarrays, Point-of-care diagnostics, Colorimetric detection, Signal enhancement, Isothermal DNA amplification, Solid-phase DNA amplification
National Category
Medical Biotechnology
Research subject
Biotechnology
Identifiers
urn:nbn:se:kth:diva-244430 (URN)978-91-7873-101-5 (ISBN)
Public defence
2019-03-22, Science for Life Laboratory, room Air & Fire, Tomtebodavägen 23A, Solna, 10:00 (English)
Opponent
Supervisors
Note

QC 20190221

Available from: 2019-02-21 Created: 2019-02-20 Last updated: 2019-02-21Bibliographically approved

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Svedberg, GustavGantelius, Jesper

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