Surface Plasmon Resonance Imaging based Multiplex Biosensor: Integration of Biomolecular Screening, Detection and Kinetics Estimation

Surface Plasmon Resonance Imaging based Multiplex Biosensor: Integration of

Biomolecular Screening, Detection and Kinetics Estimation

We present a multiplex biosensing method to simultaneously screen targets of interest in a multiple target analyte sample and to extract the binding affinities of all interactant pairs from a single sensor surface using a commercial surface plasmon resonance imaging system. For demonstration, we have prepared our sensor disk with five different ligands varying from low molecular weight antibiotics to high molecular weight human IgG, all immobilized in a microarray format. The multi-target analyte sample was prepared by mixing five antibodies where each one is highly specific for one of the immobilized ligands in a range of concentrations for kinetics estimations. The key advantage of the newly developed approach is that many different types of assays can be performed simultaneously, however, care should be taken to understand the non-specific interactions between different analytes in the sample mixture and unintended ligands, and surface regeneration behavior of different ligand types. Other advantages include reductions in experimental and analysis time, reduced costs, and flexibility since the same microarray can be used for assays with a single target analyte specific for the single ligand.

Fig. 1. (a) Schematic illustration of the immobilized spot locations. (b) Real-time iSPR image of the array of 24 immobilized spots. The numbers represent the spot numbers with respect to the molecules used: color indicates the respective color of the molecules indicated in (a). (c) Schematic illustration of the classical experiment where single target analytes are injected over the array and its measured response for the specific spot. (d) Schematic illustration of newly developed approach while injecting a multiple target analytes (each specific for different ligands immobilized) and its measured responses for all the different spots.

A detailed description can be found in the research paper published in Sensors & Actuators B - Chemical Journal “Article in Press” section on 28^th April 2010:

http://dx.doi.org/10.1016/j.snb.2010.04.015

For more information please contact Ganeshram Krishnamoorthy: g.krishnamoorthy@utwente.nl