Fabrication of carboxylated silicon nitride sensor chips for detection of antigen-antibody reaction using microfluidic reflectometric interference spectroscopy.

Yoshikazu Kurihara, Masaaki Takama, Tadanobu Sekiya, Yuka Yoshihara, Tooru Ooya, Toshifumi Takeuchi

Graduate School of Engineering, Kobe University, 1-1 Rokkodai-cho, Nada-ku, Kobe 657-8501, Japan.

Langmuir : the ACS journal of surfaces and colloids 2012 Sep 25

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In this study, we report label-free detection of alpha-fetoprotein (AFP), which has been used as a biomarker for hepatocellular carcinoma, by a microfluidic reflectometric interference spectroscopy (RIfS) system adopting a simple halogen light source and an inexpensive silicon-based sensor chip. Introduction of carboxy groups on a silicon nitride sensor chip to immobilize anti-AFP monoclonal antibody (anti-AFP) was carried out simply by immersion in aqueous solution containing triethoxysilylpropylmaleamic acid bearing a carboxy group and a silanol group. The RIfS system with the anti-AFP-immobilized sensor chip was found to give a reversible response through 100 on/off cycles using a regeneration buffer with high reproducibility (coefficient of variation (CV) = 5.7%). The limit of detection (LOD) of AFP was 100 ng mL(-1), and the measurement range spanned 3 orders of magnitude. Furthermore, the sensor chip showed no cross-reactivity with human serum albumin, Immunoglobulin G, transferrin, or fibrinogen at 100 μg mL(-1) without the use of blocking reagents such as bovine serum albumin. Consequently, the proposed RIfS system is a potentially effective tool for biomarker detection and in vitro diagnostics.

Citation

Yoshikazu Kurihara, Masaaki Takama, Tadanobu Sekiya, Yuka Yoshihara, Tooru Ooya, Toshifumi Takeuchi. Fabrication of carboxylated silicon nitride sensor chips for detection of antigen-antibody reaction using microfluidic reflectometric interference spectroscopy. Langmuir : the ACS journal of surfaces and colloids. 2012 Sep 25;28(38):13609-15