To achieve improved sensitivity in cardiac biomarker detection, a batch incubation magnetic microbead immunoassay was developed and tested on three separate human protein targets: myoglobin, heart-type fatty acid binding protein, and cardiac troponin I. A sandwich immunoassay was performed in a simple micro-centrifuge tube allowing full dispersal of the solid capture surface during incubations. Following magnetic bead capture and wash steps, samples were analyzed in the presence of a manipulated magnetic field utilizing a modified microscope slide and fluorescent inverted microscope to collect video data files. Analysis of the video data allowed for the quantitation of myoglobin, heart-type fatty acid binding protein and cardiac troponin I to levels of 360 aM, 67 fM, and 42 fM, respectively. Compared to the previous detection limit of 50 pM for myoglobin, this offers a five-fold improvement in sensitivity. This improvement in sensitivity and incorporation of additional markers, along with the small sample volumes required, suggest the potential of this platform for incorporation as a detection method in a total sample analysis device enabling multiplexed detection for the analysis of clinical samples.
Details
- Sensitive Detection of Cardiac Biomarkers Using a Magnetic Microbead Immunoassay
- Woolley, Christine (Author)
- Hayes, Mark (Author)
- Department of Chemistry and Biochemistry (Contributor)
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Digital object identifier: 10.1039/C5AY01071C
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Identifier TypeInternational standard serial numberIdentifier Value1759-9660
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Link to article as published: http://pubs.rsc.org/en/content/articlelanding/2015/ay/c5ay01071c
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Woolley, Christine, & Hayes, Mark A. (2015) Sensitive detection of cardiac biomarkers using a magnetic microbead immunoassay. ANALYTICAL METHODS. 7, 8632-8639. DOI: 10.1039/C5AY01071C