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Design of a rapid point-of-care immunoassay : Based on a single-step non-competitive immunoassay with magnetic permeability detection

Ibraimi, Filiz LU (2018)
Abstract (Swedish)

Abstract
This thesis describes the development of a one-step, specific, cost-effective and rapid in vitro point-of-care immunoassay test based on a unique magnetic immunoassay principle. The principle in question is based on a non-competitive heterogeneous immunoassay in a one-step analysis, relying on magnetic permeability detection. The heterogeneous immunoassay is performed in disposable reagent vials, containing solid phase (antibody conjugated silica microparticles) and a labelling agent (antibody conjugated superparamagnetic nanoparticles) always in suspension. The sample (antigen) is introduced into the reagent vial and mixed, forming a sandwich complex between the conjugated nano- and microparticles. The complex is sedimented to the bottom... (More)
This thesis describes the development of a one-step, specific, cost-effective and rapid in vitro point-of-care immunoassay test based on a unique magnetic immunoassay principle. The principle in question is based on a non-competitive heterogeneous immunoassay in a one-step analysis, relying on magnetic permeability detection. The heterogeneous immunoassay is performed in disposable reagent vials, containing solid phase (antibody conjugated silica microparticles) and a labelling agent (antibody conjugated superparamagnetic nanoparticles) always in suspension. The sample (antigen) is introduced into the reagent vial and mixed, forming a sandwich complex between the conjugated nano- and microparticles. The complex is sedimented to the bottom of the reagent vial, forming a pellet with increased magnetic permeability, and subsequently measured using an inductance-based magnetic permeability detector.

As proof-of-principle, an initial analyte target human C-reactive protein (CRP), a cardiac and inflammation marker in whole blood was chosen. The developed magnetic immunoassay system was characterized by a low detection limit and high accuracy. The immunoassay system was further optimized considering assay time and sample volume for better suitability in a point-of-care setting. Additionally, the two-site immunoassay was enhanced for several target analytes and sample media: human albumin, an indicator of kidney malfunction, in undiluted urine, and canine CRP, acute phase marker for identifying inflammation, in whole blood. Finally, the canine CRP assay was further developed, based on studies of the binding kinetics and reaction mechanisms, improving the sensitivity and extending the linear dynamic range, including inclusion of features suitable for point-of-care use.

In summary, the designed one step two-site heterogeneous immunoassay system, utilizing magnetic permeability based detection was shown to be a suitable and reliable method for diagnostic measurements and point-of-care use.
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author
supervisor
opponent
  • Professor Mandenius, Carl-Fredrik, Linköping University, Linköping, Sweden
organization
publishing date
type
Thesis
publication status
published
subject
keywords
Superparamagnetic nanoparticles, magnetic permeability, two-site immunoassay, point-of-care, diagnostic test, biomarkers
pages
138 pages
publisher
Department of Chemistry, Lund University
defense location
lecture hall C, Kemicentrum, Naturvetarvägen 14, Lund University, Faculty of Engineering LTH, Lund
defense date
2018-02-16 13:15
ISBN
978-91-7422-561-7
language
English
LU publication?
yes
id
bd32fb6e-b336-429a-ae7c-ef97e6983988
date added to LUP
2018-01-23 14:00:11
date last changed
2018-01-24 07:56:47
@phdthesis{bd32fb6e-b336-429a-ae7c-ef97e6983988,
  abstract     = {This thesis describes the development of a one-step, specific, cost-effective and rapid in vitro point-of-care immunoassay test based on a unique magnetic immunoassay principle. The principle in question is based on a non-competitive heterogeneous immunoassay in a one-step analysis, relying on magnetic permeability detection. The heterogeneous immunoassay is performed in disposable reagent vials, containing solid phase (antibody conjugated silica microparticles) and a labelling agent (antibody conjugated superparamagnetic nanoparticles) always in suspension. The sample (antigen) is introduced into the reagent vial and mixed, forming a sandwich complex between the conjugated nano- and microparticles. The complex is sedimented to the bottom of the reagent vial, forming a pellet with increased magnetic permeability, and subsequently measured using an inductance-based magnetic permeability detector.<br/><br/>As proof-of-principle, an initial analyte target human C-reactive protein (CRP), a cardiac and inflammation marker in whole blood was chosen. The developed magnetic immunoassay system was characterized by a low detection limit and high accuracy. The immunoassay system was further optimized considering assay time and sample volume for better suitability in a point-of-care setting. Additionally, the two-site immunoassay was enhanced for several target analytes and sample media: human albumin, an indicator of kidney malfunction, in undiluted urine, and canine CRP, acute phase marker for identifying inflammation, in whole blood. Finally, the canine CRP assay was further developed, based on studies of the binding kinetics and reaction mechanisms, improving the sensitivity and extending the linear dynamic range, including inclusion of features suitable for point-of-care use.<br/><br/>In summary, the designed one step two-site heterogeneous immunoassay system, utilizing magnetic permeability based detection was shown to be a suitable and reliable method for diagnostic measurements and point-of-care use.<br/>},
  author       = {Ibraimi, Filiz},
  isbn         = {978-91-7422-561-7 },
  keyword      = {Superparamagnetic nanoparticles, magnetic permeability, two-site immunoassay, point-of-care, diagnostic test, biomarkers},
  language     = {eng},
  pages        = {138},
  publisher    = {Department of Chemistry, Lund University},
  school       = {Lund University},
  title        = {Design of a rapid point-of-care immunoassay : Based on a single-step non-competitive immunoassay with magnetic permeability detection},
  year         = {2018},
}