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

描述（由申请人提供）：标记抗体是生物学和医学中最广泛使用的工具之一。诸如分析物免疫测定、流式细胞术和蛋白质免疫定位的方法在纯研究和临床诊断应用中被大规模使用。荧光标记抗体的发展扩大了这些技术的范围和能力，允许同时检测几个实体，同时提高整体灵敏度。然而，尽管进行了大量的技术最大化努力，但目前的荧光探针不能提供许多应用所需的多路复用能力和灵敏度。这些限制是由于i）使用可见光谱进行荧光检测和ii）所用荧光团的固有性质。 提出的研究旨在通过开发标记有新型荧光团的抗体来克服这些限制，这些荧光团在光谱的广泛未使用区域-短波红外（SWIR）（850- 1200 nm）中发射。在该区域中可以以更高的灵敏度检测荧光发射，因为与可见光区域相比，生物材料 在这些波长下显示出可忽略的自发荧光。更重要的是，可用于检测的光谱区域的加倍将大大提高多重分析的能力。待使用的荧光团是单个种类的单壁碳纳米管（SWCNT）。这些新型碳结构的荧光发射的其他独特性质将进一步有助于它们在这些应用中的有用性。单个单壁碳纳米管物种显示出尖锐的独特的发射光谱，提供更大的复用可能性比常用的可见光范围的荧光。与常用的荧光剂不同，SWCNT荧光显示没有光漂白或间歇发射（“闪烁”）。单壁碳纳米管荧光检测的灵敏度也是显著的：单个单壁碳纳米管可以在生物材料中检测到。这些单壁碳纳米管为基础的免疫探针的成功产生将促进广泛使用的临床应用和基础研究。特别地，通过同时探测多种细胞标志物检测转移性细胞的确定性将显著提高。 公共卫生相关性：荧光标记的抗体广泛用于临床应用，用于检测激素、血液蛋白、疾病生物体和癌细胞。然而，这些抗体的当前版本在检测灵敏度和可同时用于单个临床样品的数量方面具有限制。提出的实验将创造新的荧光抗体，克服这些限制，从而提高医疗诊断的质量和效率。

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.