| Antifouling surface layers for improved signal-to-noise of particle-based immunoassays. | |
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MedLine Citation:
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PMID: 19928944 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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A 10-fold improvement in the signal-to-noise (S/N) ratio of an optically encoded silica particle-based immunoassay was achieved through incorporating a protein resistant poly(ethylene glycol) (PEG) surface layer and optimizing antibody immobilization conditions. PEG was activated using 2,2,2-trifluoroethanesulfonyl chloride (tresyl) and required a minimum reaction time of 1.5 h. The activated PEG had a reactive half-life of approximately 5 h when stored in acidified dimethyl sulfoxide (DMSO). By increasing the protein incubation time and concentration, a maximum antibody loading on the particle surface of 1.6 x 10(-2) molecules per nm(2) was achieved. The assay S/N ratio was assessed using a multiplexed multicomponent optically encoded species-specific immunoassay. Encoded particles were covalently grafted or nonspecifically coated with either bovine or mouse IgG for the simultaneous detection of complementary anti-IgG "target" or uncomplementary anti-IgG "noise". The versatility and potential as a serum-based assay platform was demonstrated by immobilizing either a polyclonal antibody or an engineered single-chain variable fragment (scFv) capture probe on particles for the detection of the ovarian cancer biomarker, mesothelin (MSLN). The MLSN antigen was spiked into PBS buffer or 50% human serum. Both capture probe orientations, and media conditions showed similar low level detection limits of 5 ng/mL; however, a 40% decrease in maximum signal intensity was observed for assays run in 50% serum. |
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Authors:
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Annie Chen; Darby Kozak; Bronwyn J Battersby; Robin M Forrest; Nathalie Scholler; Nicole Urban; Matt Trau |
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Publication Detail:
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Type: Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Langmuir : the ACS journal of surfaces and colloids Volume: 25 ISSN: 1520-5827 ISO Abbreviation: Langmuir Publication Date: 2009 Dec |
Date Detail:
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Created Date: 2009-11-25 Completed Date: 2010-02-01 Revised Date: 2011-10-26 |
Medline Journal Info:
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Nlm Unique ID: 9882736 Medline TA: Langmuir Country: United States |
Other Details:
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Languages: eng Pagination: 13510-5 Citation Subset: IM |
Affiliation:
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Biomarker Research and Development Centre, Level 5 East, Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, St. Lucia, QLD 4072, Australia. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Cattle GPI-Linked Proteins Humans Immunoassay / methods* Membrane Glycoproteins / analysis Mice Nanoparticles / chemistry Polyethylene Glycols / chemistry Sulfones / chemistry |
| Grant Support | |
ID/Acronym/Agency:
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P50 CA083636-11/CA/NCI NIH HHS; P50CA083636/CA/NCI NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/GPI-Linked Proteins; 0/Membrane Glycoproteins; 0/Polyethylene Glycols; 0/Sulfones; 0/mesothelin; 1648-99-3/2,2,2-trifluoroethanesulfonyl chloride |
| Comments/Corrections | |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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