A HIGHLY SENSITIVE MICROCANTILEVER-BASED IMMUNOSENSOR ARRAY

基于微悬臂梁的高灵敏度免疫传感器阵列

• 批准号: 0617240
• 负责人: Ashok Mulchandani
• 金额: $ 100万
• 依托单位: University of California-Riverside
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-15 至 2010-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0617240&HistoricalAwards=false
• 关键词: HIGHLY; SENSITIVE; MICROCANTILEVER; BASED; IMMUNOSENSOR

0617240MulchandaniThe overall objective is to develop a novel microcantilever-based array immunosensor which is highly-sensitive, highly selective, precise, accurate, rapid, field-deployable, robust, low-cost and allows the simultaneous detection and quantification of small analytes. The proposed sensor will be based on the combination of the force measurement transduction mechanism and the receptor-ligand biological interaction recognition principle to realize the high sensitivity and selectivity. Trinitrotoluene (TNT), atrazine (ATZ), and biotin will be studied as model analytes. Single chain antibody fragment (scFv) against TNT and ATZ with low affinity and high off-rate for the respective analogs will be generated using directed evolution. The selected scFv will be immobilized on the microcantilever tip to which agarose beads functionalized with the analog will be suspended and the adhesive force measured. The introduction of the target analyte that has higher affinity and high on-rate to this system will result in the displacement of the attached bead and hence rupture of the adhesive force. The time required to rupture the linkage will be correlated to the concentration of the analyte. The knowledge gained and the sensors developed through this research will introduce novel biosensor design concepts for the applications in health care, homeland security, food monitoring and quality control, etc. The proposed effort will have impact on teaching and training of the next generation work force in the traditional areas of molecular biology and physics and the emerging multidisciplinary subject of sensors/biosensors, nanobiotechnology and microfabrication.

总体目标是开发一种新型的基于微阵列的免疫传感器，其具有高灵敏度、高选择性、精确、准确、快速、现场可部署、稳健、低成本，并且允许同时检测和定量小分析物。该传感器将基于力测量转换机理和受体-配体生物相互作用识别原理相结合，实现高灵敏度和高选择性。将三硝基甲苯（TNT）、阿特拉津（ATZ）和生物素作为模型分析物进行研究。使用定向进化将产生针对TNT和ATZ的单链抗体片段（scFv），其对相应类似物具有低亲和力和高解离速率。选择的单链抗体将固定在微悬臂梁尖端上，用类似物功能化的琼脂糖珠将悬浮在该尖端上，并测量粘附力。将具有更高亲和力和高结合速率的目标分析物引入该系统将导致附着的珠粒的位移，从而破坏粘附力。断开连接所需的时间将与分析物的浓度相关。通过本研究获得的知识和开发的传感器将为医疗保健，国土安全，食品监测和质量控制等应用引入新的生物传感器设计概念。拟议的努力将对分子生物学和物理学传统领域的下一代工作人员的教学和培训以及新兴的传感器/生物传感器多学科产生影响，纳米生物技术和微加工。