Engineered Self-Assembling Fvs for Piezoimmunosensors

用于压电免疫传感器的工程自组装 Fv

基本信息

批准号：
7848738
负责人：
XIANGQUN ZENG
金额：
$ 1.82万
依托单位：
OAKLAND UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2003
资助国家：
美国
起止时间：
2003-09-12 至 2009-10-31
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): Piezoimmunosensors (PZs) are biosensors in which antibodies are placed on a quartz crystal microbalance (QCM) to detect minute changes in mass as the antibodies bind with antigens. However, use of PZ technology in biosensors is problematic due to the complex nature of whole antibodies, which cause problems with non-specific interactions and molecule trapping. To overcome these problems, an approach in which recombinant antibody variable fragments (Fv) in self assembling monolayers (SAM) serve as the biorecognition agent is proposed. Engineered Fvs will form a "lawn" of antigen-binding sites coating the surface of the QCM, providing a sensing layer which is chemically stable, contains high numbers of binding sites, and forms a thin uniform surface. The initial study will focus on developing an a-human/goat IgG Fv- SAM to test the underlying principle. Four bioengineering methods will be used to create an Fv-SAM that possesses the necessary attributes. Electrochemistry, network impedance analysis, and Atomic Force Microscope (AFM) will be used in parallel to characterize the immobilized Fv-SAM layers for defects/pin hole structure, rigidity, orientation, stability, and surface coverage. The Fv-SAM piezoimmunosensor will be evaluated and validated for sensitivity, selectivity, linearity, stability, detection limits, and reliability in comparison to other immunoassays and/or traditional methods (GC/LC or GC/MS, LC/MS). An array of sensors specific for various antigens/toxins could be automated or combined with flow injection systems to assay different analytes in complex samples with on-line display of results. The project could lead to production of an inexpensive, easy-to-use biosensor which provides fast, precise identification of biological agents

描述（由申请人提供）：压电免疫传感器（PZS）是生物传感器，其中将抗体放在石英晶体微量平衡（QCM）上，以检测抗体与抗原结合时质量的微小变化。但是，由于整个抗体的复杂性质，在生物传感器中使用PZ技术是有问题的，这会导致非特异性相互作用和分子捕获的问题。为了克服这些问题，提出了一种方法，其中提出了自组装单层（SAM）中的重组抗体可变片段（FV）作为生物识别剂。工程的FV将形成一个“草坪”的抗原结合位点涂在QCM表面，提供化学稳定的传感层，包含大量的结合位点，并形成薄均匀的表面。最初的研究将着重于开发A-Human/Goat IgG FV- SAM来测试基本原理。四种生物工程方法将用于创建具有必要属性的FV-SAM。电化学，网络阻抗分析和原子力显微镜（AFM）将平行使用，以表征固定的FV-SAM层用于缺陷/销孔结构，刚度，方向，稳定性，稳定性和表面覆盖率。与其他免疫测定和/或传统方法（GC/LC或GC/MS，LC/MS）相比，将对FV-SAM压电传感器进行评估和验证。特定于各种抗原/毒素的传感器阵列可以自动化或与流入系统自动化，以在在线显示结果的复杂样品中分析不同的分析物。该项目可能导致生产廉价，易于使用的生物传感器，该生物传感器可快速，精确地识别生物剂