公开(公告)号：WO2007032788A2

公开(公告)日：2007-03-22

申请号：PCT/US2006/019463

申请日：2006-05-18

Applicant: SANDIA NATIONAL LABORATORIES

Inventor： HERR, Amy, Elizabeth ,  SINGH, Anup, K. ,  THROCKMORTON, Daniel, J.

IPC: B01D57/02 ,  G01N27/447

CPC classification number: G01N27/44726 ,  B82Y10/00 ,  B82Y30/00 ,  G01N27/44747

Abstract: A micro-analytical platform for performing electrophoresis-based immunoassays was developed by integrating photopolymerized cross-linked polyacrylamide gels within a microfluidic device. The microfluidic immunoassays are performed by gel electrophoretic separation and quantifying analyte concentration based upon conventional polyacrylamide gel electrophoresis (PAGE). To retain biological activity of proteins and maintain intact immune complexes, native PAGE conditions were employed. Both direct (non-competitive) and competitive immunoassay formats are demonstrated in microchips for detecting toxins and biomarkers (cytokines, c-reactive protein) in bodily fluids (serum, saliva, oral fluids). Further, a description of gradient gels fabrication is included, in an effort to describe methods we have developed for further optimization of on-chip PAGE immunoassays. The described chip-based PAGE immunoassay method enables immunoassays are fast (minutes) and require very small amounts of sample (less than a few microliters). Use of microfabricated chips as a platform enables integration, parallel assays, automation and development of portable devices.

Abstract translation: 通过将光聚合的交联聚丙烯酰胺凝胶集成在微流体装置内，开发了用于执行基于电泳的免疫测定的微分析平台。 基于常规聚丙烯酰胺凝胶电泳（PAGE），通过凝胶电泳分离和定量分析物浓度进行微流体免疫测定。 为了保留蛋白质的生物学活性并维持完整的免疫复合物，采用天然PAGE条件。 用于检测体液（血清，唾液，口腔液）中的毒素和生物标志物（细胞因子，c-反应蛋白）的微芯片中都证明了直接（非竞争性）和竞争性免疫测定形式。 此外，还包括对梯度凝胶制造的描述，以描述我们为进一步优化片上PAGE免疫测定而开发的方法。 所描述的基于芯片的PAGE免疫测定方法使免疫测定快速（分钟）并且需要非常少量的样品（少于几微升）。 使用微型芯片作为平台可实现便携式设备的集成，并行分析，自动化和开发。

公开(公告)号：WO2007032789A2

公开(公告)日：2007-03-22

申请号：PCT/US2006/019464

申请日：2006-05-18

Applicant: SANDIA NATIONAL LABORATORIES ,  HERR, Amy, Elizabeth ,  SINGH, Anup, K. ,  THROCKMORTON, Daniel, J.

Inventor： HERR, Amy, Elizabeth ,  SINGH, Anup, K. ,  THROCKMORTON, Daniel, J.

IPC: C12M1/34 ,  G01N33/558

CPC classification number: G01N27/447 ,  B82Y10/00 ,  B82Y15/00 ,  B82Y30/00 ,  G01N27/44747

Abstract: A micro-analytical platform for performing electrophoresis-based immunoassays was developed by integrating photopolymerized cross-linked polyacrylamide gels within a microfluidic device. The microfluidic immunoassays are performed by gel electrophoretic separation and quantifying analyte concentration based upon conventional polyacrylamide gel electrophoresis (PAGE). To retain biological activity of proteins and maintain intact immune complexes, native PAGE conditions were employed. Both direct (non-competitive) and competitive immunoassay formats are demonstrated in microchips for detecting toxins and biomarkers (cytokines, c-reactive protein) in bodily fluids (serum, saliva, oral fluids). Further, a description of gradient gels fabrication is included, in an effort to describe methods we have developed for further optimization of on-chip PAGE immunoassays. The described chip-based PAGE immunoassay method enables immunoassays are fast (minutes) and require very small amounts of sample (less than a few microliters). Use of microfabricated chips as a platfoπn enables integration, parallel assays, automation and development of portable devices.

Abstract translation: 通过将光聚合的交联聚丙烯酰胺凝胶整合到微流体装置内，开发了用于进行基于电泳的免疫测定的微分析平台。 通过凝胶电泳分离和定量基于常规聚丙烯酰胺凝胶电泳（PAGE）的分析物浓度来进行微流体免疫测定。 为了保持蛋白质的生物活性并保持完整的免疫复合物，采用天然PAGE条件。 用于检测体液（血清，唾液，口服液）中的毒素和生物标志物（细胞因子，c反应蛋白）的微芯片都展示了直接（非竞争性）和竞争性免疫测定形式。 此外，包括梯度凝胶制造的描述，以努力描述我们为进一步优化片上PAGE免疫测定而开发的方法。 所述基于片段的PAGE免疫测定方法使得免疫测定是快速（分钟）并且需要非常少量的样品（少于几微升）。 使用微加工芯片作为平台，可实现便携式设备的集成，并行测定，自动化和开发。