Microfluidic tectonics platform: A colorimetric, disposable botulinum toxin enzyme-linked immunosorbent assay system
Microfluidic tectonics platform: A colorimetric, disposable botulinum toxin enzyme-linked immunosorbent assay system
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DOI：
10.1002/elps.200405888
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发表时间：
2004-06-01
期刊：
影响因子：
2.9
通讯作者：
Beebe, DJ
中科院分区：
文献类型：
--
作者：
Moorthy, J;Mensing, GA;Beebe, DJ
讨论了实现集成微流控器件的制造平台。该平台允许以特别的方式创建用于多步测定的特定微系统，因为执行测定步骤的组件可以经由原位制造在装置内的任何位置处创建。该平台被用来创建一个原型微系统，用于检测肉毒杆菌神经毒素直接从全血。在该装置上进行了过滤、混合和与试剂孵育等样品制备等工艺步骤。由单体、交联剂和光引发剂组成的预聚物混合物制备了各种微流控组件，如通道网络、阀和多孔过滤器。为了检测类毒素，将生物素化的抗体固定在链霉亲和素官能化的琼脂糖凝胶珠上。将凝胶珠引入装置中并用作读数。碱性磷酸酶（在二抗上）和底物之间的酶促反应产生不溶性有色沉淀物，该沉淀物包被珠粒，从而使读数肉眼可见。使用便携式酶联免疫吸附测定（ELISA）系统可以从全血中检测到临床相关量的毒素。多层可以实现有效的空间利用和创建一个三维（3-D）混沌混合器。此外，外部材料如膜可作为组件并入装置中。进行这些步骤所需的各个组件进行了表征，并讨论了它们的相互兼容性。
A fabrication platform for realizing integrated microfluidic devices is discussed. The platform allows for creating specific microsystems for multistep assays in an ad hoc manner as the components that perform the assay steps can be created at any location inside the device via in situ fabrication. The platform was utilized to create a prototype microsystem for detecting botulinum neurotoxin directly from whole blood. Process steps such as sample preparation by filtration, mixing and incubation with reagents was carried out on the device. Various microfluidic components such as channel network, valves and porous filter were fabricated from prepolymer mixture consisting of monomer, cross-linker and a photoinitiator. For detection of the toxoid, biotinylated antibodies were immobilized on streptavidin-functionalized agarose gel beads. The gel beads were introduced into the device and were used as readouts. Enzymatic reaction between alkaline phosphatase (on secondary antibody) and substrate produced an insoluble, colored precipitate that coated the beads thus making the readout visible to the naked eye. Clinically relevant amounts of the toxin can be detected from whole blood using the portable enzyme-linked immunosorbent assay (ELISA) system. Multiple layers can be realized for effective space utilization and creating a three-dimensional (3-D) chaotic mixer. In addition, external materials such as membranes can be incorporated into the device as components. Individual components that were necessary to perform these steps were characterized, and their mutual compatibility is also discussed.