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Antibody immobilization on to polystyrene substrate—on-chip immunoassay for horse IgG based on fluorescence

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Abstract

A simple microfluidic immunoassay card was developed based on polystyrene (PS) substrate for the detection of horse IgG, an inexpensive model analyte using fluorescence microscope. The primary antibody was captured onto the PS based on covalent bonding via a self-assembled monolayer (SAM) of thiol to pattern the surface chemistry on a gold-coated PS. The immunosensor chip layers were fabricated from sheets by CO2 laser ablation. The functionalized PS surfaces after each step were characterized by contact angle measurement, X-ray photoelectron spectroscopy (XPS), and atomic force microscopy (AFM). After the antibody–antigen interaction as a sandwich immunoassay with a fluorescein isothiocyanate (FITC)-conjugated secondary antibody, the intensity of fluorescence was measured on-chip to determine the concentration of the target analyte. The present immunosensor chip showed a linear response range for horse IgG between 1 μg/ml and 80 μg/ml (r = 0.971, n = 3). The detection limit was found to be 0.71 μg/ml. The developed microfluidic system can be extended for various applications including medical diagnostics, microarray detection and observing protein–protein interactions.

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Acknowledgements

This work was supported by Biomedical Engineering Research Council of Singapore under the Singapore-University of Washington Alliance program. F. Darain would like to thank Ms. Fang Xiaoqin from School of Electrical and Electronic Engineering for her help to make the design to cut the layers for microfluidic devices.

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Authors and Affiliations

  1. Biomedical Engineering Research Centre, Nanyang Technological University, Singapore, 639553, Singapore

    Farzana Darain & Kai Ling Gan

  2. Photonics Research Centre, School of Electrical and Electronic Engineering, Nanyang Technological University, Singapore, 639798, Singapore

    Swee Chuan Tjin

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  1. Farzana Darain
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  2. Kai Ling Gan
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  3. Swee Chuan Tjin
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Correspondence to Farzana Darain.

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Darain, F., Gan, K.L. & Tjin, S.C. Antibody immobilization on to polystyrene substrate—on-chip immunoassay for horse IgG based on fluorescence. Biomed Microdevices 11, 653–661 (2009). https://doi.org/10.1007/s10544-008-9275-3

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