Novel probes for expanding immunobased detection into the short wave infrared spe

用于将基于免疫的检测扩展到短波红外光谱的新型探针

• 批准号: 8302855
• 负责人: Kathleen Mary Beckingham
• 金额: $ 20.29万
• 依托单位: RICE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-09 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8302855
• 关键词: Antibodies; Basic Science; Biocompatible Materials; Biological; Biology; Biomedical Engineering; Blinking; Blood; Blood Proteins; Cancer Detection; Cancerous; Carbon; Cell model; Cell surface; Cells; Chemistry; Clinical; Collaborations; Detection; Development; Diagnosis; Diagnostic; Disease; Drosophila genus; Evaluation; Family; Flow Cytometry; Fluorescence; Fluorescent Antibody Technique; Fluorescent Probes; Generations; Healthcare; Hormones; Image; Immunoassay; Individual; Infectious Agent; Institutes; Label; Laboratories; Larva; Link; Measurement; Medical; Medicine; Methods; Monitor; Nanotubes; Newborn Infant; Optics; Organism; Performance; Photobleaching; Play; Polystyrenes; Property; Proteins; Research; Role; Sampling; Screening procedure; Short Waves; Sorting - Cell Movement; Structure; Suspension substance; Suspensions; System; Techniques; Testing; Time; Tissue Sample; Tissues; Width; base; brain tissue; clinical Diagnosis; clinical application; comparative; cytokine; fluorescence imaging; fluorophore; high sensitivity probe; improved; instrument; novel; research study; single walled carbon nanotube; tool

DESCRIPTION (provided by applicant): Tagged antibodies are one of most widely used tools in biology and medicine. Methods such as analyte immunoassays, flow cytometry, and protein immunolocalization are used on an enormous scale in both pure research and clinical, diagnostic applications. The development of fluorescently labeled antibodies has extended the range and power of these techniques by permitting simultaneous detection of several entities while at the same time increasing overall sensitivity. However, despite intense technical maximization efforts, current fluorescent probes do not provide the multiplexing capacity and sensitivity desired for many applications. These limitations result from i) the use of the visible spectrum for fluorescence detection and ii) the inherent properties of the fluorophores used. The studies proposed are aimed at overcoming these limitations by developing antibodies tagged with novel fluorophores that emit in a broad, unused region of the spectrum - the shortwave infrared (SWIR) (850- 1200 nm). Fluorescence emission can be detected with greater sensitivity in this region because, in contrast to the visible region, biological materials show negligible autofluorescence at these wavelengths. More importantly, doubling of the spectral region that can be used for detection will greatly enhance capacity for multiplexed analysis. The fluorophores to be used are individual species of single walled carbon nanotubes (SWCNTs). Additional unique properties of the fluorescent emission from these novel carbon structures will contribute further to their usefulness in these applications. Individual SWCNT species show sharp distinct emission spectra, providing greater multiplexing possibilities than the commonly used visible-range fluors. Unlike the commonly used fluors, SWCNT fluorescence shows no photobleaching or intermittent emission ("blinking"). The sensitivity for SWCNT fluorescence detection is also remarkable: individual SWCNTs can be detected in biological material. The successful generation of these SWCNT-based immunoprobes will enhance widely used clinical applications and basic research. In particular, the certainty of detection of metastatic cells by simultaneous probing for multiple cell markers will be significantly improved. PUBLIC HEALTH RELEVANCE: Fluorescently labeled antibodies are widely used in clinical applications for detection of hormones, blood proteins, disease organisms, and cancerous cells. However, the current versions of these antibodies have limitations in terms of sensitivity of detection and the number that can be used simultaneously on a single clinical sample. The experiments proposed will create novel fluorescent antibodies that will overcome these limitations and thus improve the quality and efficiency of medical diagnoses.

描述（由申请人提供）：标记的抗体是生物学和医学中最广泛使用的工具之一。在纯研究和临床，诊断应用中，诸如分析物免疫测定，流式细胞仪和蛋白质免疫定位等方法均在巨大的范围内使用。荧光标记的抗体的开发通过允许同时检测几个实体，同时提高总体灵敏度，从而扩大了这些技术的范围和功率。然而，尽管技术最大化的努力强烈，但当前的荧光探针并未为许多应用提供多重能力和灵敏度。这些局限性源于i）可见光谱用于荧光检测以及ii）所用荧光团的固有特性。 提出的研究旨在通过开发带有新型荧光团标记的抗体来克服这些局限性，这些抗体在光谱的广阔未使用区域-Shortwave红外（SWIR）（SWIR）（850-1200 nm）。在该区域中，可以以更高的敏感性检测荧光发射，因为与可见区域相反，生物材料 在这些波长处显示可忽略的自动荧光。更重要的是，可用于检测的光谱区域的加倍将大大提高多路复用分析的能力。要使用的荧光团是单个壁碳纳米管（SWCNT）的单个物种。这些新型碳结构的荧光发射的其他独特特性将进一步有助于它们在这些应用中的用处。单个SWCNT物种显示出明显的不同发射光谱，比常用的可见范围荧光提供了更大的多重可能性。与常用的荧光不同，SWCNT荧光没有光漂白或间歇发射（“闪烁”）。 SWCNT荧光检测的敏感性也很引人注目：可以在生物材料中检测到单个SWCNT。这些基于SWCNT的免疫探针的成功产生将增强广泛使用的临床应用和基础研究。特别是，通过同时探测多个细胞标记的转移细胞检测的确定性将得到显着改善。 公共卫生相关性：荧光标记的抗体被广泛用于检测激素，血蛋白，疾病生物和癌细胞的临床应用中。但是，这些抗体的当前版本在检测敏感性方面存在局限性，并且可以同时在单个临床样本上使用的数量。提出的实验将创建新型的荧光抗体，以克服这些局限性，从而提高医疗诊断的质量和效率。