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

描述(申请人提供)：标记抗体是生物学和医学中使用最广泛的工具之一。分析物免疫分析、流式细胞术和蛋白质免疫定位等方法在纯研究和临床诊断应用中都得到了广泛的应用。荧光标记抗体的发展扩展了这些技术的范围和力量，允许同时检测几个实体，同时提高了总体灵敏度。然而，尽管进行了密集的技术最大化努力，目前的荧光探针并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以及ii)所使用的荧光团的固有性质造成的。建议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一个广泛的、未使用的区域--短波红外(SWIR)(850-1200 nm)--发射。在这个区域可以更灵敏地探测到荧光发射，因为与可见光区域相比，生物材料 在这些波长显示可以忽略不计的自发荧光。更重要的是，可用于检测的光谱区域加倍将大大增强多路分析的能力。将使用的荧光团是单个种类的单壁碳纳米管(SWCNTs)。来自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应用。单个单壁碳纳米管物种显示出清晰的发射光谱，提供了比常用的可见光范围荧光更大的多路复用可能性。与通常使用的荧光不同，SWCNT荧光不会出现光漂白或间歇性发射(“闪烁”)。SWCNT荧光检测的灵敏度也很高：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促进其广泛的临床应用和基础研究。特别是，通过同时探测多个细胞标志物来检测转移细胞的确定性将显著提高。 公共卫生相关性：荧光标记抗体广泛用于检测激素、血液蛋白、疾病生物体和癌细胞的临床应用。然而，当前版本的这些抗体在检测灵敏度和可在单个临床样本上同时使用的数量方面存在局限性。拟议的实验将创造出新的荧光抗体，以克服这些限制，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.