Homogeneous immunoassay based on aggregation of antibody-functionalized gold nanoparticles coupled with light scattering detection.
A universal platform of homogeneous noncompetitive immunoassay, using human immunoglobulin (IgG) as a model analyte, has been developed. The assay is based on aggregation of antibody-functionalized gold nanoparticles directed by the immunoreaction coupled with light scattering detection with a common spectrofluorimeter. In phosphate buffer (pH 7.0) solution, the light scattering intensity of the gold nanoparticles functionalized with goat-anti-human IgG can be greatly enhanced by addition of the human IgG. Based on this phenomenon, a wide dynamic range of 0.05-10 microg ml(-1) for determination of human IgG can be obtained, and the detection limit can reach 10 ng ml(-1). The proposed immunoassay can be accomplished in a homogeneous solution with one-step operation within 10 min and has been successfully applied to the determination of human IgG in serum samples, in which the results are well consistent with those of the enzyme-linked immunosorbent assay (ELISA), indicating its high selectivity and practicality. Therefore, the gold nanoparticle-based light scattering method can be used as a model to establish the general methods for protein assay in the fields of molecular biology and clinical diagnostics.
Du
Baoan
B
College of Chemistry and Environmental Science, Hebei University, Baoding 072002, PR China.
Li
Zhengping
Z
Cheng
Yongqiang
Y
D000818:Animals; D000941:Antigens / Q000276:immunology; D006041:Goats / Q000276:immunology; D006046:Gold / Q000737:chemistry*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007118:Immunoassay / Q000379:methods*; D007074:Immunoglobulin G / Q000032:analysis* / Q000737:chemistry* / Q000276:immunology; D007185:Incubators; D008027:Light; D053768:Metal Nanoparticles / Q000737:chemistry*; D009994:Osmolar Concentration; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence; D013997:Time Factors
D000941:Antigens; D007074:Immunoglobulin G; D006046:Gold