Homogeneous immunoassay for soy protein determination in food samples using gold nanoparticles as labels and light scattering detection.
A homogeneous aggregation immunoassay involving the use of gold nanoparticles (AuNPs) and light scattering detection is described for soy protein determination in food samples. AuNPs act as enhancers of the precipitate that appears when the antigen-antibody complex is formed. The AuNPs-antibody conjugate has been synthesized by physical adsorption of polyclonal anti-soy protein antibodies onto the surface of commercial AuNPs with a nominal diameter of 20nm. The direct assay is based on the reaction of the conjugate with soy protein, which reaches the equilibrium in about 10min, and the measurement of the light scattering intensity at 530nm, which is proportional to the analyte concentration. The dynamic range of the calibration graph is 0.2-20microgm L(-1) and the detection limit value is 65ngm L(-1). The precision, expressed as relative standard deviation, has been assayed at two different concentrations, 0.2 and 1microgm L(-1), giving values ranging from 4.7 to 5.9%. The interference of other proteins has been assayed. The usefulness of this method has been shown by its application to the analysis of fruit juice and "nonmilk yoghourt" samples. The results obtained with the proposed method are similar to those obtained by using a commercial ELISA kit, but the assay time is significantly shorter and the detection limit was about 10 times lower. A recovery study has been also performed, giving values in the range of 84.0-119.3%.
Sánchez-Martínez
M L
ML
Department of Analytical Chemistry, University of Córdoba, Campus of Rabanales, Marie-Curie Annex Building, 14071 Córdoba, Spain.
Aguilar-Caballos
M P
MP
Gómez-Hens
A
A
D005504:Food Analysis; D005638:Fruit / Q000737:chemistry; D006046:Gold / Q000737:chemistry*; D007118:Immunoassay / Q000379:methods*; D053768:Metal Nanoparticles / Q000737:chemistry*; D015203:Reproducibility of Results; D012542:Scattering, Radiation; D030262:Soybean Proteins / Q000032:analysis*
D030262:Soybean Proteins; D006046:Gold