Biosensor for Rapid Detection of Prion

用于快速检测朊病毒的生物传感器

• 批准号: 6934289
• 负责人: THOMAS G HAZEL
• 金额: $ 11.03万
• 依托单位: INNOVATIVE BIOSENSORS, INC.
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-08-01 至 2007-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6934289
• 关键词: B lymphocyte; antigen antibody reaction; bioluminescence; biosensor device; biotechnology; calcium indicator; cell line; diagnosis design /evaluation; diagnostic tests; hybridomas; luciferin monooxygenase; monoclonal antibody; polymerase chain reaction; portable biomedical equipment; prions; rapid diagnosis

DESCRIPTION (provided by applicant): Creutzfeldt-Jakob Disease and Bovine Spongiform Encephalopathy are transmissible, neurodegenerative and fatal prion diseases of humans and cattle, respectively. There are no commercially available, completely reliable diagnostic tests for use before the onset of symptoms. The only reliable tests in use are performed post mortem or via brain biopsy and involve histological examination. For these reasons, there is a great need for better diagnostics for these prion diseases. This application proposes to develop a rapid, extremely sensitive diagnostic test for prion based on the CANARY biosensor technology developed at MIT and an antibody that specifically recognizes the scrapie form of the prion protein in cattle and humans. The CANARY technology is composed of B-lymphocytes that have been genetically engineered to express an antibody of interest on the cell surface and calcium sensitive protein in the cell's cytosol. The interaction of the antigen with the membrane bound antibody cause the engineered biosensors to emit light, which is detectable by a small, portable luminometer. This approach allows specific testing of analytes at previously unachievable levels of speed and sensitivity. The specific tasks for Phase I include: 1) Obtaining sequences of the light and heavy chains of the variable region of the anti-prion monoclonal antibody by RT-PCR. 2) Construction of these sequences into a well characterized two vector antibody expression system. 3) Generation of a stable cell line that expresses the recombinant antibody on its surface and a calcium sensitive bioluminescent protein in the cell's cytosol that responds to the prion antibody epitope.

描述（由申请人提供）：克鲁特兹菲尔特 - 贾科布疾病和牛海绵状脑病分别是可传播的，神经退行性和致命的人类和牛的致命疾病。在症状发作之前，没有商业可用的可靠诊断测试。使用的唯一可靠测试是在验尸后或通过脑活检进行的，并涉及组织学检查。由于这些原因，对于这些病毒疾病，需要更好的诊断。 该应用程序建议基于MIT开发的金丝雀生物传感器技术和一种专门识别牛和人类中prion蛋白的crapie形式的抗体，开发针对prion的快速，极其敏感的诊断测试。金丝雀技术由B淋巴细胞组成，这些细胞经过基因设计，用于在细胞表面表达抗体，并在细胞的细胞质中表达钙敏感蛋白。抗原与膜结合抗体的相互作用导致工程生物传感器发出光，可通过小的便携式发光计检测到光。这种方法允许在先前无法实现的速度和灵敏度水平上对分析物进行特定测试。 第一阶段的特定任务包括：1）通过RT-PCR获得抗prion抗原单克隆抗体的可变区域的光和重链的序列。 2）将这些序列的构造成两个表征的两个载体抗体表达系统。 3）生成稳定的细胞系，该细胞系在其表面上表达重组抗体，并在细胞的胞质溶胶中钙敏感的生物发光蛋白，从而对prion抗体表位做出反应。