Departamento de Química, Instituto de Reconocimiento Molecular y Desarrollo Tecnológico, Universidad Politécnica de Valencia, Camino de Vera s/n, 46071 Valencia, Spain.
Anal Chem. 2010 Mar 1;82(5):1954-63. doi: 10.1021/ac902706t.
An amino-modified polycarbonate surface of compact discs is used to link haptens covalently and directly as an alternative to the classic protein-hapten conjugate adsorption coating strategy employed in immunoassays. The modified surface maintains its physical and optical properties, and a standard disk drive can then read the assay results. Advantages are evaluated, such as the use of a broader spectrum of coupling media including organic solvents that are inappropriate for proteins but necessary for some water-insoluble haptens and the bypassing of the synthesis and purification for protein conjugates. As proof of concept, competitive microimmunoassays were developed for chlorpyrifos, atrazine, and 2-(2,4,5-trichlorophenoxy)propionic acid (2,4,5-TP), in microarray format, obtaining detection limits of 37.2, 8.1, and 76 ng/L, respectively. The sensitivity was 1 order of magnitude better than that obtained for all the studied systems using hapten-protein conjugates adsorbed on polystyrene enzyme-linked immunosorbent assay (ELISA) plates and polycarbonate surfaces. Further, the influence of hapten structure and presentation on molecular recognition pattern is discussed. To our knowledge, this is the first time that microarray and compact disc technologies converge with this particular hapten immobilization mode. The great potential of the approach is demonstrated through the high-throughput capability of the disc in a range of analytical applications, as well as the inherent advantages of compact disc reading technology.
光盘的氨基修饰聚碳酸酯表面被用于将半抗原共价且直接连接，作为免疫分析中经典的蛋白-半抗原缀合吸附涂层策略的替代方法。修饰后的表面保持其物理和光学性质，然后标准的磁盘驱动器可以读取分析结果。评估了其优点，例如可以使用更广泛的偶联介质，包括不适合蛋白质但对于一些水溶性差的半抗原是必要的有机溶剂，并且可以绕过蛋白缀合物的合成和纯化。作为概念验证，以微阵列格式开发了用于氯吡磷、莠去津和 2-(2,4,5-三氯苯氧基)丙酸（2,4,5-TP）的竞争性微免疫分析，分别获得 37.2、8.1 和 76 ng/L 的检测限。与使用吸附在聚苯乙烯酶联免疫吸附测定（ELISA）板和聚碳酸酯表面上的半抗原-蛋白缀合物的所有研究体系相比，灵敏度提高了 1 个数量级。此外，还讨论了半抗原结构和呈现方式对半抗原分子识别模式的影响。据我们所知，这是微阵列和光盘技术首次与这种特殊的半抗原固定化模式相结合。该方法的巨大潜力通过光盘在各种分析应用中的高通量能力以及光盘读取技术的固有优势得到了证明。
