DETECTION OF ANTI-HUMAN SERUM ALBUMIN ANTIBODY USING SURFACE PLASMON RESONANCE BIOSENSOR

摘要

The field of surface plasmon resonace(SPR) sensor has attracted much attention in recent years as a tool for studying macromolecular interactions since it is a "direct optical sensing technique" not requiring any labels . In this paper, BIAcore 1000 instrument based on SPR technology was used as an immunobiosensor to detect a series of goat anti-human serum albumin (anti-HSA) solutions with different concentrations. The methodology relies on the covalent immobilization of one of the macromolecules involved in the interaction under study onto the sensing surface, a hydrophilic carboxymethylated dextran-modified gold surface. Immobilization of the ligand is achieved by activation of the carboxyl groups of the dextran to produce N-hydroxysuccinimide esters (NHS-esters). Primary amines of the ligand react with the activated carboxyls to produce amide bonds. The analyte of interest is then injected across the immobilized ligand surface and the interaction of soluble analyte is observed directly, in real time, as a change in the SPR signal. The interacting analyte can then be eluted (desorbed) so that the "regenerated" surface may be used for subsequent analyte binding measurements.