State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuchang, Wuhan 430072, PR China.
Talanta. 2009 Nov 15;80(1):407-10. doi: 10.1016/j.talanta.2009.06.044. Epub 2009 Jun 27.
An indirect inhibitive surface plasmon resonance (SPR) immunoassay was developed for the microcystins (MCs) detection. The bioconjugate of MC-LR and bovine serum albumin (BSA) was immobilized on a CM5 sensor chip. A serial premixture of MC-LR standards (or samples) and monoclonal antibody (mAb) were injected over the functional sensor surface, and the subsequent specific immunoreaction was monitored on the BIAcore 3000 biosensor and generated a signal with an increasing intensity in response to the decreasing MCs concentration. The developed SPR immunoassay has a wide quantitative range in 1-100 microg L(-1). Although not as sensitive as conventional enzyme-linked immunosorbent assay (ELISA), the SPR biosensor offered unique advantages: (1) the sensor chip could be reusable without any significant loss in its binding activity after 50 assay-regeneration cycles, (2) one single assay could be accomplished in 50 min (including 30-min preincubation and 20-min BIAcore analysis), and (3) this method did not require multiple steps. The SPR biosensor was also used to detect MCs in environmental samples, and the results compared well with those obtained by ELISA. We conclude that the SPR biosensor offers outstanding advantages for the MCs detection and may be further developed as a field-portable sensor for real-time monitoring of MCs on site in the near future.
建立了一种用于检测微囊藻毒素（MCs）的间接抑制表面等离子体共振（SPR）免疫分析方法。将 MC-LR 和牛血清白蛋白（BSA）的生物缀合物固定在 CM5 传感器芯片上。将一系列 MC-LR 标准品（或样品）和单克隆抗体（mAb）混合物注入功能传感器表面，随后在 BIAcore 3000 生物传感器上监测到特定的免疫反应，并根据 MCs 浓度的降低生成强度不断增加的信号。开发的 SPR 免疫分析方法在 1-100 μg L(-1) 范围内具有广泛的定量范围。尽管不如传统的酶联免疫吸附测定（ELISA）灵敏，但 SPR 生物传感器具有独特的优势：（1）在 50 次测定-再生循环后，传感器芯片可以重复使用，而不会显著损失其结合活性；（2）单次测定可以在 50 分钟内完成（包括 30 分钟的预孵育和 20 分钟的 BIAcore 分析）；（3）该方法不需要多个步骤。SPR 生物传感器还用于检测环境样品中的 MCs，结果与 ELISA 获得的结果非常吻合。我们得出结论，SPR 生物传感器在 MCs 检测方面具有突出的优势，并且可能在不久的将来进一步开发为现场实时监测 MCs 的便携式传感器。
