Document Detail

Antifouling surface layers for improved signal-to-noise of particle-based immunoassays.
MedLine Citation:
PMID:  19928944     Owner:  NLM     Status:  MEDLINE    
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.
Annie Chen; Darby Kozak; Bronwyn J Battersby; Robin M Forrest; Nathalie Scholler; Nicole Urban; Matt Trau
Related Documents :
3955814 - Direct measurement of creatine kinase-mb activity in serum after extraction with a mono...
7074904 - Theoretical and mathematical conditions for use of linearization methods for saturation...
8903514 - Monoclonal antibodies that specifically recognize crystals of dinitrobenzene.
1487514 - Compared stability of sepharose-based immunoadsorbents prepared by various activation m...
1756224 - Non-polarized cell surface expression of hla-a,b,c and hla-dr antigens in graves' thyro...
15001314 - Protein a/g dependent elisa a promising diagnostic tool in lyme disease seroprevalence ...
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:  2014-09-08    
Medline Journal Info:
Nlm Unique ID:  9882736     Medline TA:  Langmuir     Country:  United States    
Other Details:
Languages:  eng     Pagination:  13510-5     Citation Subset:  IM    
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
GPI-Linked Proteins
Immunoassay / methods*
Membrane Glycoproteins / analysis
Nanoparticles / chemistry
Polyethylene Glycols / chemistry
Sulfones / chemistry
Grant Support
P50 CA083636/CA/NCI NIH HHS; P50 CA083636-11/CA/NCI NIH HHS; P50CA083636/CA/NCI NIH HHS
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

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

Previous Document:  Thermally Induced Radical Hydrosilylation for Synthesis of C18 HPLC phases from Highly Condensed SiH...
Next Document:  Polymer Migration among Phospholipid Liposomes.