Effect of poly(ethylene glycol), tetramethylammonium hydroxide, and other surfactants on enhancing performance in a latex particle immunoassay of C-reactive protein.
The influence of a variety and combination of both ionic surfactants and different chain lengths of the polyelectrolyte poly(ethylene glycol) (PEG) on the performance characteristics (with particular reference to signal response) of a homogeneous, latex agglutination immunoassay was investigated. The test analyte was human serum C-reactive protein (CRP), and the antibody reagent consisted of a sheep polyclonal anti-CRP IgG fraction covalently coupled to 50-nm-sized latex including a glycine-capped chloromethylstyrene shell. The amount and rate of immunoagglutination was monitored turbidimetrically after sample addition. It was found that 2.5 mmol/L concentrations of the small cationic surfactant tetramethylammonium hydroxide (TMH), when present alone, substantially increased both reaction rates and sensitivity in the lower clinical ranges of CRP concentration when compared to normally used assay conditions containing PEG and the anionic detergent Gafac. The nonspecific binding (NSB) was also found to be unchanged. Evidence is presented that the TMH enhances the actual antibody-antigen interaction as opposed to the known effects of other surfactants in immunocomplex dissociation or in maintenance of colloidal stability. We suggest that the enhancement seen with TMH could be an alternative to PEG and may provide a new means of further extending detection limits. The utility of this type of immunoassay technology could therefore be increased whenever clinically required.
Holownia
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P
Department of Clinical Biochemistry, St. Bartholomews and Royal London Hospital's School of Medicine and Dentistry, UK. p.g.holownia@mds.qmw.ac.uk
Perez-Amodio
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S
Price
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CP
D000838:Anions / Q000737:chemistry; D002097:C-Reactive Protein / Q000032:analysis*; D002412:Cations / Q000737:chemistry; D006801:Humans; D007118:Immunoassay / Q000379:methods; D007841:Latex Fixation Tests / Q000379:methods*; D011092:Polyethylene Glycols / Q000737:chemistry*; D000644:Quaternary Ammonium Compounds / Q000737:chemistry*; D013501:Surface-Active Agents / Q000737:chemistry*
D000838:Anions; D002412:Cations; D000644:Quaternary Ammonium Compounds; D013501:Surface-Active Agents; D011092:Polyethylene Glycols; D002097:C-Reactive Protein; C027917:tetramethylammonium