A New Method for Immobilizing Antibody Probes in Immunochips Based on DNA Hybridization

Abstract

Based on complementary DNA strands hybridization,a method for immobilizing antibodies on immunochips was established.In this method,each kind of antibodies was conjugated to a specific sequence of oligonucleotide,and then was immobilized onto a complementary-oligonucleotide coated solid surface via base pairing.The method was first testified on magnetic beads surface.An immunochip against transgenic Bt Cry1Ac protein was further developed based on this in mobilization method.The immobilization outcome was analyzed using Cy3 labeled secondary antibody,and gradient concentrations of Bt Cry1Ac protein were tested on the developed chip.The result showed the chip using this immobilization method achieved good specificity,even distribution of antibody probes,little non-specific adsorption,and a detection limit of 0.01-0.05 μg/L.Finally,the immunochip could be regenerated through dissociating the two DNA strands.