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Gold nanoparticle-based fluorescence immunoassay for malaria antigen detection.(Report)

Byline: Bassem S. S. Guirgis (1,2), Claudia Sa e Cunha (3), Ines Gomes (1,3), Miguel Cavadas (1), Isabel Silva (1), Goncalo Doria (4), Gregory L. Blatch (5,6), Pedro V. Baptista (4), Eulalia Pereira (7), Hassan M. E. Azzazy (2), Maria M. Mota (3), Miguel Prudencio (3), Ricardo Franco (1) Keywords: M... Full description

Journal Title: Analytical and Bioanalytical Chemistry Jan, 2012, Vol.402(2), p.1019(9)
Main Author: Guirgis, Bassem S. S.
Other Authors: Sa E Cunha, Claudia , Gomes, Ines , Cavadas, Miguel , Silva, Isabel , Doria, Goncalo , Blatch, Gregory L. , Baptista, Pedro V. , Pereira, Eulalia , Azzazy, Hassan M. E. , Mota, Maria M. , Prudencio, Miguel , Franco, Ricardo
Format: Electronic Article Electronic Article
Language: English
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Quelle: Cengage Learning, Inc.
ID: ISSN: 1618-2642
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title: Gold nanoparticle-based fluorescence immunoassay for malaria antigen detection.(Report)
format: Article
creator:
  • Guirgis, Bassem S. S.
  • Sa E Cunha, Claudia
  • Gomes, Ines
  • Cavadas, Miguel
  • Silva, Isabel
  • Doria, Goncalo
  • Blatch, Gregory L.
  • Baptista, Pedro V.
  • Pereira, Eulalia
  • Azzazy, Hassan M. E.
  • Mota, Maria M.
  • Prudencio, Miguel
  • Franco, Ricardo
subjects:
  • Skilled Labor -- Analysis
  • Heat Shock Proteins -- Analysis
  • Adsorption -- Analysis
  • Anopheles -- Analysis
  • Malaria -- Analysis
  • Antigens -- Analysis
  • Fluorescence -- Analysis
  • Plasmodium Falciparum -- Analysis
  • Monoclonal Antibodies -- Analysis
  • Protein Binding -- Analysis
  • Universities And Colleges -- Analysis
ispartof: Analytical and Bioanalytical Chemistry, Jan, 2012, Vol.402(2), p.1019(9)
description: Byline: Bassem S. S. Guirgis (1,2), Claudia Sa e Cunha (3), Ines Gomes (1,3), Miguel Cavadas (1), Isabel Silva (1), Goncalo Doria (4), Gregory L. Blatch (5,6), Pedro V. Baptista (4), Eulalia Pereira (7), Hassan M. E. Azzazy (2), Maria M. Mota (3), Miguel Prudencio (3), Ricardo Franco (1) Keywords: Malaria diagnosis; Gold nanoparticles; Nanodiagnosis; Heat shock protein; Fluorescence detection; Immunoassay; Plasmodium Abstract: The development of rapid detection assays for malaria diagnostics is an area of intensive research, as the traditional microscopic analysis of blood smears is cumbersome and requires skilled personnel. Here, we describe a simple and sensitive immunoassay that successfully detects malaria antigens in infected blood cultures. This homogeneous assay is based on the fluorescence quenching of cyanine 3B (Cy3B)-labeled recombinant Plasmodium falciparum heat shock protein 70 (PfHsp70) upon binding to gold nanoparticles (AuNPs) functionalized with an anti-Hsp70 monoclonal antibody. Upon competition with the free antigen, the Cy3B-labeled recombinant PfHsp70 is released to solution resulting in an increase of fluorescence intensity. Two types of AuNP-antibody conjugates were used as probes, one obtained by electrostatic adsorption of the antibody on AuNPs surface and the other by covalent bonding using protein cross-linking agents. In comparison with cross-linked antibodies, electrostatic adsorption of the antibodies to the AuNPs surfaces generated conjugates with increased activity and linearity of response, within a range of antigen concentration from 8.2 to 23.8 ug.mL.sup.-1. The estimated LOD for the assay is 2.4 ug.mL.sup.-1 and the LOQ is 7.3 ug.mL.sup.-1. The fluorescence immunoassay was successfully applied to the detection of antigen in malaria-infected human blood cultures at a 3% parasitemia level, and is assumed to detect parasite densities as low as 1,000 parasites.uL.sup.-1. Figure A simple and sensitive competitive immunoassay based on fluorescence-quenching by gold nanoparticles, successfully detected malaria antigens in blood cultures infected with 1,000 parasites.uL.sup.-1. Author Affiliation: (1) REQUIMTE, Departamento de Quimica, Faculdade de Ciencias e Tecnologia, Universidade Nova de Lisboa, 2829-516, Caparica, Portugal (2) Department of Chemistry, SSE 1194 and Yousef Jameel Science & Technology Research Center, The American University in Cairo, AUC Avenue, PO Box 74, New Cairo, 11835, Egypt (3) Instituto de Medicina Mol
language: English
source: Cengage Learning, Inc.
identifier: ISSN: 1618-2642
fulltext: fulltext
issn:
  • 1618-2642
  • 16182642
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titleGold nanoparticle-based fluorescence immunoassay for malaria antigen detection.(Report)
creatorGuirgis, Bassem S. S. ; Sa E Cunha, Claudia ; Gomes, Ines ; Cavadas, Miguel ; Silva, Isabel ; Doria, Goncalo ; Blatch, Gregory L. ; Baptista, Pedro V. ; Pereira, Eulalia ; Azzazy, Hassan M. E. ; Mota, Maria M. ; Prudencio, Miguel ; Franco, Ricardo
ispartofAnalytical and Bioanalytical Chemistry, Jan, 2012, Vol.402(2), p.1019(9)
identifierISSN: 1618-2642
subjectSkilled Labor -- Analysis ; Heat Shock Proteins -- Analysis ; Adsorption -- Analysis ; Anopheles -- Analysis ; Malaria -- Analysis ; Antigens -- Analysis ; Fluorescence -- Analysis ; Plasmodium Falciparum -- Analysis ; Monoclonal Antibodies -- Analysis ; Protein Binding -- Analysis ; Universities And Colleges -- Analysis
descriptionByline: Bassem S. S. Guirgis (1,2), Claudia Sa e Cunha (3), Ines Gomes (1,3), Miguel Cavadas (1), Isabel Silva (1), Goncalo Doria (4), Gregory L. Blatch (5,6), Pedro V. Baptista (4), Eulalia Pereira (7), Hassan M. E. Azzazy (2), Maria M. Mota (3), Miguel Prudencio (3), Ricardo Franco (1) Keywords: Malaria diagnosis; Gold nanoparticles; Nanodiagnosis; Heat shock protein; Fluorescence detection; Immunoassay; Plasmodium Abstract: The development of rapid detection assays for malaria diagnostics is an area of intensive research, as the traditional microscopic analysis of blood smears is cumbersome and requires skilled personnel. Here, we describe a simple and sensitive immunoassay that successfully detects malaria antigens in infected blood cultures. This homogeneous assay is based on the fluorescence quenching of cyanine 3B (Cy3B)-labeled recombinant Plasmodium falciparum heat shock protein 70 (PfHsp70) upon binding to gold nanoparticles (AuNPs) functionalized with an anti-Hsp70 monoclonal antibody. Upon competition with the free antigen, the Cy3B-labeled recombinant PfHsp70 is released to solution resulting in an increase of fluorescence intensity. Two types of AuNP-antibody conjugates were used as probes, one obtained by electrostatic adsorption of the antibody on AuNPs surface and the other by covalent bonding using protein cross-linking agents. In comparison with cross-linked antibodies, electrostatic adsorption of the antibodies to the AuNPs surfaces generated conjugates with increased activity and linearity of response, within a range of antigen concentration from 8.2 to 23.8 ug.mL.sup.-1. The estimated LOD for the assay is 2.4 ug.mL.sup.-1 and the LOQ is 7.3 ug.mL.sup.-1. The fluorescence immunoassay was successfully applied to the detection of antigen in malaria-infected human blood cultures at a 3% parasitemia level, and is assumed to detect parasite densities as low as 1,000 parasites.uL.sup.-1. Figure A simple and sensitive competitive immunoassay based on fluorescence-quenching by gold nanoparticles, successfully detected malaria antigens in blood cultures infected with 1,000 parasites.uL.sup.-1. Author Affiliation: (1) REQUIMTE, Departamento de Quimica, Faculdade de Ciencias e Tecnologia, Universidade Nova de Lisboa, 2829-516, Caparica, Portugal (2) Department of Chemistry, SSE 1194 and Yousef Jameel Science & Technology Research Center, The American University in Cairo, AUC Avenue, PO Box 74, New Cairo, 11835, Egypt (3) Instituto de Medicina Molecular, Faculdade de Medicina da Universidade de Lisboa, 1649-028, Lisbon, Portugal (4) CIGMH, Departamento de Ciencias da Vida, Faculdade de Ciencias e Tecnologia, Universidade Nova de Lisboa, 2829-516, Caparica, Portugal (5) Biomedical Biotechnology Research Unit, Department of Biochemistry, Microbiology and Biotechnology, Rhodes University, Grahamstown, 6140, South Africa (6) School of Biomedical and Health Sciences, Victoria University, Melbourne, VIC, 8001, Australia (7) REQUIMTE, Departamento de Quimica e Bioquimica, Faculdade de Ciencias, Universidade do Porto, 4169-007, Porto, Portugal Article History: Registration Date: 09/10/2011 Received Date: 01/08/2011 Accepted Date: 09/10/2011 Online Date: 17/11/2011 Article note: Bassem S. S. Guirgis and Miguel Cavadas contributed equally to this work. Electronic supplementary material The online version of this article (doi: 10.1007/s00216-011-5489-y) contains supplementary material, which is available to authorized users.
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titleGold nanoparticle-based fluorescence immunoassay for malaria antigen detection.(Report)
descriptionByline: Bassem S. S. Guirgis (1,2), Claudia Sa e Cunha (3), Ines Gomes (1,3), Miguel Cavadas (1), Isabel Silva (1), Goncalo Doria (4), Gregory L. Blatch (5,6), Pedro V. Baptista (4), Eulalia Pereira (7), Hassan M. E. Azzazy (2), Maria M. Mota (3), Miguel Prudencio (3), Ricardo Franco (1) Keywords: Malaria diagnosis; Gold nanoparticles; Nanodiagnosis; Heat shock protein; Fluorescence detection; Immunoassay; Plasmodium Abstract: The development of rapid detection assays for malaria diagnostics is an area of intensive research, as the traditional microscopic analysis of blood smears is cumbersome and requires skilled personnel. Here, we describe a simple and sensitive immunoassay that successfully detects malaria antigens in infected blood cultures. This homogeneous assay is based on the fluorescence quenching of cyanine 3B (Cy3B)-labeled recombinant Plasmodium falciparum heat shock protein 70 (PfHsp70) upon binding to gold nanoparticles (AuNPs) functionalized with an anti-Hsp70 monoclonal antibody. Upon competition with the free antigen, the Cy3B-labeled recombinant PfHsp70 is released to solution resulting in an increase of fluorescence intensity. Two types of AuNP-antibody conjugates were used as probes, one obtained by electrostatic adsorption of the antibody on AuNPs surface and the other by covalent bonding using protein cross-linking agents. In comparison with cross-linked antibodies, electrostatic adsorption of the antibodies to the AuNPs surfaces generated conjugates with increased activity and linearity of response, within a range of antigen concentration from 8.2 to 23.8 ug.mL.sup.-1. The estimated LOD for the assay is 2.4 ug.mL.sup.-1 and the LOQ is 7.3 ug.mL.sup.-1. The fluorescence immunoassay was successfully applied to the detection of antigen in malaria-infected human blood cultures at a 3% parasitemia level, and is assumed to detect parasite densities as low as 1,000 parasites.uL.sup.-1. Figure A simple and sensitive competitive immunoassay based on fluorescence-quenching by gold nanoparticles, successfully detected malaria antigens in blood cultures infected with 1,000 parasites.uL.sup.-1. Author Affiliation: (1) REQUIMTE, Departamento de Quimica, Faculdade de Ciencias e Tecnologia, Universidade Nova de Lisboa, 2829-516, Caparica, Portugal (2) Department of Chemistry, SSE 1194 and Yousef Jameel Science & Technology Research Center, The American University in Cairo, AUC Avenue, PO Box 74, New Cairo, 11835, Egypt (3) Instituto de Medicina Molecular, Faculdade de Medicina da Universidade de Lisboa, 1649-028, Lisbon, Portugal (4) CIGMH, Departamento de Ciencias da Vida, Faculdade de Ciencias e Tecnologia, Universidade Nova de Lisboa, 2829-516, Caparica, Portugal (5) Biomedical Biotechnology Research Unit, Department of Biochemistry, Microbiology and Biotechnology, Rhodes University, Grahamstown, 6140, South Africa (6) School of Biomedical and Health Sciences, Victoria University, Melbourne, VIC, 8001, Australia (7) REQUIMTE, Departamento de Quimica e Bioquimica, Faculdade de Ciencias, Universidade do Porto, 4169-007, Porto, Portugal Article History: Registration Date: 09/10/2011 Received Date: 01/08/2011 Accepted Date: 09/10/2011 Online Date: 17/11/2011 Article note: Bassem S. S. Guirgis and Miguel Cavadas contributed equally to this work. Electronic supplementary material The online version of this article (doi: 10.1007/s00216-011-5489-y) contains supplementary material, which is available to authorized users.
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abstractByline: Bassem S. S. Guirgis (1,2), Claudia Sa e Cunha (3), Ines Gomes (1,3), Miguel Cavadas (1), Isabel Silva (1), Goncalo Doria (4), Gregory L. Blatch (5,6), Pedro V. Baptista (4), Eulalia Pereira (7), Hassan M. E. Azzazy (2), Maria M. Mota (3), Miguel Prudencio (3), Ricardo Franco (1) Keywords: Malaria diagnosis; Gold nanoparticles; Nanodiagnosis; Heat shock protein; Fluorescence detection; Immunoassay; Plasmodium Abstract: The development of rapid detection assays for malaria diagnostics is an area of intensive research, as the traditional microscopic analysis of blood smears is cumbersome and requires skilled personnel. Here, we describe a simple and sensitive immunoassay that successfully detects malaria antigens in infected blood cultures. This homogeneous assay is based on the fluorescence quenching of cyanine 3B (Cy3B)-labeled recombinant Plasmodium falciparum heat shock protein 70 (PfHsp70) upon binding to gold nanoparticles (AuNPs) functionalized with an anti-Hsp70 monoclonal antibody. Upon competition with the free antigen, the Cy3B-labeled recombinant PfHsp70 is released to solution resulting in an increase of fluorescence intensity. Two types of AuNP-antibody conjugates were used as probes, one obtained by electrostatic adsorption of the antibody on AuNPs surface and the other by covalent bonding using protein cross-linking agents. In comparison with cross-linked antibodies, electrostatic adsorption of the antibodies to the AuNPs surfaces generated conjugates with increased activity and linearity of response, within a range of antigen concentration from 8.2 to 23.8 ug.mL.sup.-1. The estimated LOD for the assay is 2.4 ug.mL.sup.-1 and the LOQ is 7.3 ug.mL.sup.-1. The fluorescence immunoassay was successfully applied to the detection of antigen in malaria-infected human blood cultures at a 3% parasitemia level, and is assumed to detect parasite densities as low as 1,000 parasites.uL.sup.-1. Figure A simple and sensitive competitive immunoassay based on fluorescence-quenching by gold nanoparticles, successfully detected malaria antigens in blood cultures infected with 1,000 parasites.uL.sup.-1. Author Affiliation: (1) REQUIMTE, Departamento de Quimica, Faculdade de Ciencias e Tecnologia, Universidade Nova de Lisboa, 2829-516, Caparica, Portugal (2) Department of Chemistry, SSE 1194 and Yousef Jameel Science & Technology Research Center, The American University in Cairo, AUC Avenue, PO Box 74, New Cairo, 11835, Egypt (3) Instituto de Medicina Molecular, Faculdade de Medicina da Universidade de Lisboa, 1649-028, Lisbon, Portugal (4) CIGMH, Departamento de Ciencias da Vida, Faculdade de Ciencias e Tecnologia, Universidade Nova de Lisboa, 2829-516, Caparica, Portugal (5) Biomedical Biotechnology Research Unit, Department of Biochemistry, Microbiology and Biotechnology, Rhodes University, Grahamstown, 6140, South Africa (6) School of Biomedical and Health Sciences, Victoria University, Melbourne, VIC, 8001, Australia (7) REQUIMTE, Departamento de Quimica e Bioquimica, Faculdade de Ciencias, Universidade do Porto, 4169-007, Porto, Portugal Article History: Registration Date: 09/10/2011 Received Date: 01/08/2011 Accepted Date: 09/10/2011 Online Date: 17/11/2011 Article note: Bassem S. S. Guirgis and Miguel Cavadas contributed equally to this work. Electronic supplementary material The online version of this article (doi: 10.1007/s00216-011-5489-y) contains supplementary material, which is available to authorized users.
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