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

被引:43
作者
Darain, Farzana [1 ]
Gan, Kai Ling [1 ]
Tjin, Swee Chuan [2 ]
机构
[1] Nanyang Technol Univ, Biomed Engn Res Ctr, Singapore 639553, Singapore
[2] Nanyang Technol Univ, Photon Res Ctr, Sch Elect & Elect Engn, Singapore 639798, Singapore
关键词
Polystyrene; Covalent bonding; Microfluidic card; Fluorescence; Horse IgG; SELF-ASSEMBLED MONOLAYERS; MICROFLUIDIC DEVICE; SURFACE MODIFICATION; POLYMER; IMMUNOSENSOR; ASSAY; POLY(DIMETHYLSILOXANE); SPECTROSCOPY; FILMS;
D O I
10.1007/s10544-008-9275-3
中图分类号
R318 [生物医学工程];
学科分类号
0831 ;
摘要
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 mu g/ml and 80 mu g/ml (r = 0.971, n = 3). The detection limit was found to be 0.71 mu g/ml. The developed microfluidic system can be extended for various applications including medical diagnostics, microarray detection and observing protein-protein interactions.
引用
收藏
页码:653 / 661
页数:9
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