文化大學機構典藏 CCUR:Item 987654321/20534
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    题名: Surface plasmon resonance-based immunosensor with oriented immobilized antibody fragments on a mixed self-assembled monolayer for the determination of staphylococcal enterotoxin B
    作者: Tsai, WC (Tsai, Wen-Chi)
    Pai, PJR (Pai, Pi-Ju Rini)
    贡献者: 生物科技研究所
    关键词: Surface plasmon resonance
    Mixed self-assembled monolayers
    Antibody fragments
    Staphylococcal enterotoxin B
    Immunosensor
    日期: 2009
    上传时间: 2011-11-30 14:29:42 (UTC+8)
    摘要: An immunosensor based on surface plasmon resonance (SPR) with a mixed self-assembled monolayer (SAM) was developed to determine staphylococcal enterotoxin B (SEB). The SAM on a gold surface was fabricated by adsorbing a mixture of 16-mercapto-1-hexadecanoic acid (16-MHA) and hexanethiol at various molar ratios. Initially, full-length anti-SEB was randomly immobilized onto the SAM to form the immunosensing surface. Through optimization of surface functionalization and anti-SEB immobilization, the SPR sensors can be applied to the determination of SEB in a linear range of 0.01 similar to 1.0 mu g.mL(-1). Furthermore, a smaller antibody fragment (F(ab)') was generated and immobilized randomly (via amino groups) or in an oriented manner (via -SH groups) to form the immunosensing surface. The oriented immobilization of F(ab)' led to a 50% increase in the antigen binding efficiency compared to randomly immobilized covalent F(ab') fragments. The resulting calibration curve showed higher sensitivity. In addition, the specificity and applicability of the proposed immunosensor to milk samples were also demonstrated. Furthermore, the sensor can be regenerated using 0.1 M HCl, and 70% of the initial response was maintained over 3 cycles.
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