School of Material Science and Engineering, Nanyang Technology University , Nanyang Avenue, 639798, Singapore.
Anal Chem. 2016 Jun 21;88(12):6441-9. doi: 10.1021/acs.analchem.6b01034. Epub 2016 Jun 9.
Commercially available systems that provide cost-effective, fast, simple, and portable solutions for health and environmental applications are few despite advancements in bioassays and biosensor research. We have developed a new system based on stacked membranes, each layer with a specific function. Samples were added onto the bottom-most layer, and as each layer becomes wet, the analyte pushes through to the next membrane layers. During migration, the analyte attaches with the corresponding antibody, itself conjugated with horseradish peroxidase (HRP) to produce a measurable signal. To prevent false positive results, blocking layer membranes are added to stop unbound antibodies from reaching the top membrane. Thus, only analyte/antibody-HRP complex will generate a signal. In order to prove this concept, Escherichia coli was used as the target analyte. After optimization, our immunoassay sensitivity was adjusted to 100 cells mL(-1). Different environmental water sources were also tested to demonstrate the sensitivity and specificity of our proposed stacked bioassay. Simplicity, low price, sensitivity, and modularity (capability to change to any target analyte) make this idea very promising for future commercialization.
尽管生物测定法和生物传感器研究取得了进展，但能够为健康和环境应用提供经济高效、快速、简单和便携解决方案的商业现成系统仍然很少。我们已经开发出一种基于堆叠膜的新系统，每层都具有特定的功能。将样品添加到底层，当每层变湿时，分析物就会推到下一层膜。在迁移过程中，分析物与相应的抗体结合，抗体本身与辣根过氧化物酶（HRP）缀合以产生可测量的信号。为了防止假阳性结果，添加了阻挡层膜以阻止未结合的抗体到达顶层膜。因此，只有分析物/抗体-HRP 复合物会产生信号。为了验证这一概念，我们使用大肠杆菌作为目标分析物。经过优化，我们的免疫分析灵敏度调整为 100 个细胞 mL(-1)。还测试了不同的环境水源，以证明我们提出的堆叠生物测定法的灵敏度和特异性。简单、低价、灵敏度和模块化（能够针对任何目标分析物进行更改）使这个想法非常有前途，有望实现未来的商业化。
