Design and performances of immunoassay based on SPR biosensor with magnetic microbeads

A surface plasmon resonance (SPR) biosensor system was developed for immunoassay, based on the conjugates of magnetic microbeads coupling with antibody which could be trapped on the Au film firmly due to the magnetic force. The magnetic microbeads were used as the solid support for the heat shock protein 70 (Hsp 70) antibody and antibody immobilized magnetic microbeads were utilized instead of the single antibody for the determination of Hsp 70. Since the magnetic bead is coated with dextran, the antibodies and some specific biomolecular receptors can be immobilized using a variety of chemical reactions. Compared to traditional antibody immobilization on the sensing film, there is not a covalent link between the Au film and the antibody. There is a great advantage in that sensor can be stripped and reused, and the same chemistry used to derivative dextran-coated SPR sensors can be used for the magnetic bead-coated sensors. The sensing layer was formed well. Different dilution ratios (v/v) of the conjugates result in different detectable ranges. When the dilution ratios of the conjugate are 1:10 and 1:5, the lowest concentrations of Hsp 70 that can be detected are 1.50 and 0.30 μg ml−1, respectively.