Nanotube Antibody Arrays for Profiling Circulating Diseased Cells

描述（由申请人提供）：用于对细针抽吸物、活检、全血、唾液和尿液等易于获取的身体来源中的少量细胞进行分子表征的灵敏和定量技术可能对生命科学和临床实践产生重大影响。目前用于癌症生物标志物测试的实验室程序需要收集体液和抽吸物，将这些材料转移到实验室进行测试，周转时间在一天到几天的范围内。这有时可能导致取样细胞的表型/凋亡变化。该项目将开发一种纳米管抗体生物传感器阵列，用于快速检测血液中的循环癌细胞，可以在15分钟内直接识别癌细胞上的表面标记。该项目将努力实现以下具体目标：1.检测血液中BT474、MCF 7、SKBR 3和MCF 10A癌细胞中的分子表面受体，即IGF 1 R、Her 2和EpCAM。创建表达水平和恶性度图，以在几分钟内快速分析血液中的循环肿瘤细胞。2.使用纳米针探针来研究受体与抗体的结合力，以进行非特异性和特异性相互作用，从而基于相互作用力创建表达水平和恶性度图。3.证明我们的生物传感器具有与ELISA等标准免疫测定相当的检测灵敏度和特异性。来自乳腺癌患者的血液将使用标准技术和我们的纳米管生物传感器技术进行测试，这些数据的统计相关性将有助于标准化这项技术，以供未来测试。这些目标的成功实现将导致开发一种快速且非常便宜的测试，该测试使用非常少量的血液（微升或更少）在几分钟内检测患者血液循环中的肿瘤细胞。这种廉价的测试肯定具有很强的商业潜力，因为我们计划在本提案期间验证概念后将产品商业化。 公共卫生相关性：该项目将开发纳米管抗体生物传感器阵列，用于在几分钟内快速分析血液中的循环癌细胞。基于传感器阵列，将开发一种用于检测和分析血液中循环乳腺癌细胞的设备。这项新技术将与标准免疫测定法如ELISA进行比较。

DESCRIPTION (provided by applicant): Sensitive and quantitative technologies for molecular characterization of scant cells in easily accessible bodily sources such as fine needle aspirates, biopsies, whole blood, saliva and urine can potentially have significant impacts in life sciences and clinical practice. Current laboratory procedures used for cancer biomarker testing require collection of the body fluids and aspirates, transfer of those materials to the laboratory for testing with turn-around-times in the range of a day to several days. This sometimes could lead to phenotypic/apoptotic changes of sampled cells. This project will develop a nanotube-antibody biosensor arrays for rapid detection of circulating cancer cells in blood that can identify surface markers directly on cancer cells within 15 minutes. The project will seek to accomplish the following specific aims: 1. Detection of molecular surface receptors namely IGF1R, Her2, and EpCAM in BT474, MCF7, SKBR3 and MCF10A cancer cells in blood. Creation of expression level and malignancy maps to rapidly profile the circulating tumor cells in blood within minutes. 2. Use nanoneedle probes to investigate the binding forces of receptors to antibodies for non-specific and specific interactions to create an expression level and malignancy maps based on interaction forces. 3. Demonstrate that our biosensors have the detection sensitivities and specificities that is comparable to standard immunoassays such as ELISA. Blood from breast cancer patients will be tested using both standard technique and our nanotube-biosensor technique and statistical correlation of these data will help standardize this technique for future testing. The successful accomplishment of these aims will lead to developing a quick and very inexpensive test that uses a very small quantity of blood (micro-liter or less) to detect tumor cells on a patient's blood circulation in a matter of a few minutes. This inexpensive test definitely has a strong commercial potential as we have plans for commercialization of the products after proofing the concept during the period of this proposal. PUBLIC HEALTH RELEVANCE: This project will develop nanotube-antibody biosensor arrays for rapid profiling of circulating cancer cells in blood within minutes. Based on the sensor arrays, a device will be developed that will be used in detecting and profiling circulating breast cancer cells in blood. This new technology will be compared against standard immunoassays such as ELISA.