Document Detail


Antifouling surface layers for improved signal-to-noise of particle-based immunoassays.
MedLine Citation:
PMID:  19928944     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Annie Chen; Darby Kozak; Bronwyn J Battersby; Robin M Forrest; Nathalie Scholler; Nicole Urban; Matt Trau
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Langmuir : the ACS journal of surfaces and colloids     Volume:  25     ISSN:  1520-5827     ISO Abbreviation:  Langmuir     Publication Date:  2009 Dec 
Date Detail:
Created Date:  2009-11-25     Completed Date:  2010-02-01     Revised Date:  2013-05-31    
Medline Journal Info:
Nlm Unique ID:  9882736     Medline TA:  Langmuir     Country:  United States    
Other Details:
Languages:  eng     Pagination:  13510-5     Citation Subset:  IM    
Affiliation:
Biomarker Research and Development Centre, Level 5 East, Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, St. Lucia, QLD 4072, Australia.
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MeSH Terms
Descriptor/Qualifier:
Animals
Cattle
GPI-Linked Proteins
Humans
Immunoassay / methods*
Membrane Glycoproteins / analysis
Mice
Nanoparticles / chemistry
Polyethylene Glycols / chemistry
Sulfones / chemistry
Grant Support
ID/Acronym/Agency:
P50 CA083636-11/CA/NCI NIH HHS; P50CA083636/CA/NCI NIH HHS
Chemical
Reg. No./Substance:
0/GPI-Linked Proteins; 0/Membrane Glycoproteins; 0/Polyethylene Glycols; 0/Sulfones; 0/mesothelin; 1648-99-3/2,2,2-trifluoroethanesulfonyl chloride
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