Competitive magnetic immunoassay for protein detection in thin channels

Abstract

Functional magnetic nanoparticles are prepared and characterized for protein detection in a magnetic separation channel. This detection method is based on a competitive immunoassay of magnetic separation in thin channels using functional magnetic nanoparticles. We used protein A–IgG complex to demonstrate the feasibility. Free IgG and fixed number of IgG-labeled microparticles were used to compete for limited sites of protein A on the magnetic nanoparticles. Several experimental parameters were investigated for protein detection. The deposited percentages of IgG-labeled microparticles at various concentrations of free IgG were determined and used as a reference plot. The IgG concentration in a sample was deduced and determined based on the reference plot using the deposited percentage of IgG-labeled microparticles from the sample. The linear range of IgG detection was from 5.0 × 10 −8 to 1.0 × 10 −11 M. The detection limit was 3.69 × 10 −12 M. The running time was less than 10 min. Selectivities were higher than 92% and the relative errors were less than 7%. The IgG concentration of serum was determined to be 3.6 mg ml −1. This measurement differed by 8.3% from the ELISA measurement. The recoveries of IgG spiked in serum were found to be higher than 94%. This method can provide simple, fast, and selective analysis for protein detection and other immunoassay-related applications.