High-throughput Biological-assays Via Single Molecule Labeling and Detection

通过单分子标记和检测进行高通量生物测定

• 批准号: 9876651
• 负责人: David Schultz
• 金额: $ 6.15万
• 依托单位: University of California-San Diego
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-10-01 至 2003-09-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9876651&HistoricalAwards=false
• 关键词: throughput; Biological; assays; Via; Single

ABSTRACT9876651David A. SchultzUniversity of California, San DiegoHigh-throughput Biological -assays Via Single Molecule Labeling and DetectionThis grant supports the development of a new ultrasensitive optical assay for biological macromolecules. Either protein or nucleic acid target molecules will be detected at the level of 1 femtogram. This level of sensitivity will allow the identification and quantification of the chosen macromolecule in a single cell. The sensitivity, speed, and accuracy of these assays will be optimized using a real-time in-vitro studies.The reporter molecule for such an assay must be robust and nano-sized yet still have a very large light-scattering cross-section. Immmunolabelled plasmon resonant particles (PRPs) are ideal for this purpose. PRPs are small (40-80 nanometer) metal particles that scatter visible light elastically with extraordinary efficiency when excited at their plasmon resonance frequency. They are inexpensive, non-toxic, and are available in a variety of colors. The light scattered from PRPs under typical white light illumination is so large that each PRP is individually visible. At appropriate concentration, the actual number of PRPs in a field of view is simply the number of particles bound to the target macromolecule plus any particles that are non-specifically bound. An image of such a field of PRPs can be recorded using a CCD camera within milliseconds. This will allow for the rapid and ultrasensitive detection of the target macromolecules in the cell. It is anticipated that commercialized versions of the PRP immunoassay will allow for low cost detection of a wide range of important molecules in the cell. Such an immunoassay should complement or replace standard ELISA or radio-immunoassays for cases where the concentration of the target molecules if very low and target amplification is not beneficial or practical.

摘要9876651DAVID A.加利福尼亚州的Schultzuniversity，圣地亚哥高发的生物学 - 通过单分子标记和检测来检测这一赠款支持开发用于生物大分子生物学大分子的新超敏感光学测定。 将在1 fert图的水平上检测到蛋白质或核酸靶标分子。 这种敏感性将允许单个细胞中所选的大分子的识别和定量。 这些测定法的敏感性，速度和准确性将通过实时的视频研究进行优化。这种测定的记者分子必须坚固且纳米尺寸，但仍然具有非常大的光片段横截面。 Immmunolabelled等离子体谐振颗粒（PRP）是理想的选择。 PRP是小的（40-80纳米）金属颗粒，当在其等离子体共振频率上激发时，以极其效率散射可见光。 它们是廉价，无毒的，并且有多种颜色。 从典型的白光照明下从PRP中散布的光线如此之大，以至于每个PRP单独可见。 在适当的浓度下，视野中的实际PRP数仅仅是与目标大分子的颗粒数，以及任何非特异性结合的粒子。 可以使用毫秒内的CCD摄像头记录这样的PRP领域的图像。 这将允许对细胞中靶标大分子的快速和超敏感性检测。 预计PRP免疫测定版的商业化版本将允许对细胞中广泛的重要分子进行低成本检测。 这样的免疫测定应在目标分子的浓度（如果非常低且目标放大是无益或实用的情况下）的情况下补充或替换标准ELISA或放射免疫测定。