Virus-Coated Layer-by-Layer Colloids as a Multiplex Suspension Array for the Detection and Quantification of Virus-Specific Antibodies
Suspension array technology has surpassed ELISA for automated, simultaneous detection and quantification of soluble biomarkers such as virus-specific antibodies. We describe assays in which antigens are attached to a lipid bilayer surrounding color-coded particles. We used layer-by-layer technology...
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| Veröffentlicht in: | Clinical chemistry (Baltimore, Md.) Md.), 2006-08, Vol.52 (8), p.1575-1583 |
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| creator | Toellner, Lars Fischlechner, Martin Ferko, Boris Grabherr, Reingard Maria Donath, Edwin |
| description | Suspension array technology has surpassed ELISA for automated, simultaneous detection and quantification of soluble biomarkers such as virus-specific antibodies. We describe assays in which antigens are attached to a lipid bilayer surrounding color-coded particles.
We used layer-by-layer technology to establish a multiplex suspension array with distinguishable microbeads coated with authentic viral surfaces to catch and quantify virus-specific antibodies in a flow cytometric analysis. Antigenic surfaces were generated by chimeric and wild-type baculoviruses plus 2 different influenza A virus subtypes fused to a lipid bilayer surrounding distinctly colored particles. Specificity of binding of chosen antibodies and sera was detected by immunofluorescence. Results of multiplex analysis were compared with results of ELISA.
Titrations of virus-specific antibodies in the multiplex suspension array demonstrated specific binding to the viral surface proteins. The multiplex suspension array gave positive results for up to log 5-diluted primary antibodies with an approximately 5- to 10-fold reduced dynamic range compared with the respective ELISA.
The bead-based multiplex suspension array is customizable and easy to establish. By displaying native influenza A virus surfaces and recombinant HIV-1 epitopes, the new assay provides a tool for the detection of major viral infections in humans. |
| doi_str_mv | 10.1373/clinchem.2005.065789 |
| format | Article |
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We used layer-by-layer technology to establish a multiplex suspension array with distinguishable microbeads coated with authentic viral surfaces to catch and quantify virus-specific antibodies in a flow cytometric analysis. Antigenic surfaces were generated by chimeric and wild-type baculoviruses plus 2 different influenza A virus subtypes fused to a lipid bilayer surrounding distinctly colored particles. Specificity of binding of chosen antibodies and sera was detected by immunofluorescence. Results of multiplex analysis were compared with results of ELISA.
Titrations of virus-specific antibodies in the multiplex suspension array demonstrated specific binding to the viral surface proteins. The multiplex suspension array gave positive results for up to log 5-diluted primary antibodies with an approximately 5- to 10-fold reduced dynamic range compared with the respective ELISA.
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Titrations of virus-specific antibodies in the multiplex suspension array demonstrated specific binding to the viral surface proteins. The multiplex suspension array gave positive results for up to log 5-diluted primary antibodies with an approximately 5- to 10-fold reduced dynamic range compared with the respective ELISA.
The bead-based multiplex suspension array is customizable and easy to establish. By displaying native influenza A virus surfaces and recombinant HIV-1 epitopes, the new assay provides a tool for the detection of major viral infections in humans.</description><subject>Analytical, structural and metabolic biochemistry</subject><subject>Antibodies, Viral - blood</subject><subject>Antigens</subject><subject>Antigens, Viral - chemistry</subject><subject>Baculoviridae - chemistry</subject><subject>Baculoviridae - genetics</subject><subject>Baculovirus</subject><subject>Biological and medical sciences</subject><subject>Colloids</subject><subject>Diagnostics</subject><subject>Enzyme-Linked Immunosorbent Assay</subject><subject>Flow Cytometry</subject><subject>Fluorescent Antibody Technique</subject><subject>Fundamental and applied biological sciences. 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Academic</collection><jtitle>Clinical chemistry (Baltimore, Md.)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Toellner, Lars</au><au>Fischlechner, Martin</au><au>Ferko, Boris</au><au>Grabherr, Reingard Maria</au><au>Donath, Edwin</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Virus-Coated Layer-by-Layer Colloids as a Multiplex Suspension Array for the Detection and Quantification of Virus-Specific Antibodies</atitle><jtitle>Clinical chemistry (Baltimore, Md.)</jtitle><addtitle>Clin Chem</addtitle><date>2006-08-01</date><risdate>2006</risdate><volume>52</volume><issue>8</issue><spage>1575</spage><epage>1583</epage><pages>1575-1583</pages><issn>0009-9147</issn><eissn>1530-8561</eissn><coden>CLCHAU</coden><abstract>Suspension array technology has surpassed ELISA for automated, simultaneous detection and quantification of soluble biomarkers such as virus-specific antibodies. We describe assays in which antigens are attached to a lipid bilayer surrounding color-coded particles.
We used layer-by-layer technology to establish a multiplex suspension array with distinguishable microbeads coated with authentic viral surfaces to catch and quantify virus-specific antibodies in a flow cytometric analysis. Antigenic surfaces were generated by chimeric and wild-type baculoviruses plus 2 different influenza A virus subtypes fused to a lipid bilayer surrounding distinctly colored particles. Specificity of binding of chosen antibodies and sera was detected by immunofluorescence. Results of multiplex analysis were compared with results of ELISA.
Titrations of virus-specific antibodies in the multiplex suspension array demonstrated specific binding to the viral surface proteins. The multiplex suspension array gave positive results for up to log 5-diluted primary antibodies with an approximately 5- to 10-fold reduced dynamic range compared with the respective ELISA.
The bead-based multiplex suspension array is customizable and easy to establish. By displaying native influenza A virus surfaces and recombinant HIV-1 epitopes, the new assay provides a tool for the detection of major viral infections in humans.</abstract><cop>Washington, DC</cop><pub>Am Assoc Clin Chem</pub><pmid>16728470</pmid><doi>10.1373/clinchem.2005.065789</doi><tpages>9</tpages><oa>free_for_read</oa></addata></record> |
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| source | MEDLINE; Oxford University Press Journals All Titles (1996-Current) |
| subjects | Analytical, structural and metabolic biochemistry Antibodies, Viral - blood Antigens Antigens, Viral - chemistry Baculoviridae - chemistry Baculoviridae - genetics Baculovirus Biological and medical sciences Colloids Diagnostics Enzyme-Linked Immunosorbent Assay Flow Cytometry Fluorescent Antibody Technique Fundamental and applied biological sciences. Psychology Hemagglutinin Glycoproteins, Influenza Virus - analysis Human immunodeficiency virus 1 Humans Immunoassay Immunoassay - methods Influenza A virus Influenza A virus - chemistry Investigative techniques, diagnostic techniques (general aspects) Lipid Bilayers - chemistry Medical sciences Microspheres Viral Fusion Proteins - analysis Virology |
| title | Virus-Coated Layer-by-Layer Colloids as a Multiplex Suspension Array for the Detection and Quantification of Virus-Specific Antibodies |
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