New Class of Bright, Sharp, Tunable Near-Infrared Fluorophores for Flow Cytometry

用于流式细胞术的新型明亮、锐利、可调谐近红外荧光团

• 批准号: 9234452
• 负责人: James Bruce Pitner
• 金额: $ 75.68万
• 依托单位: NIRVANA SCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-06-15 至 2018-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9234452
• 关键词: Adjuvant; Agreement; Autoimmune Diseases; Basic Science; Biological Assay; Biological Markers; Cell Count; Cells; Chemicals; Color; Communicable Diseases; Core Facility; Development; Disease; Dyes; Effectiveness; Emerging Communicable Diseases; Evaluation; Exhibits; Experimental Designs; Family; Financial compensation; Flow Cytometry; Fluorescent Dyes; Goals; HIV; HIV vaccine; Hypersensitivity; Immune; Immune response; Immune system; Immunologics; Immunology; Immunophenotyping; Immunotherapy; Label; Laboratories; Lasers; Malaria; Mathematics; Mediating; Methods; Modernization; Modification; National Institute of Allergy and Infectious Disease; North Carolina; Performance; Phase; Polymers; Preparation; Procedures; Production; Protocols documentation; Reagent; Research; Resolution; Route; Safety; Sampling; Science; Sensitivity and Specificity; Small Business Technology Transfer Research; Source; Techniques; Technology; Testing; Transplantation; Tuberculosis; Ultraviolet Rays; Universities; Vaccine Research; Validation; Water; Width; antibody conjugate; bacteriochlorin; clinical diagnostics; design; experimental study; fluorophore; improved; member; novel; novel strategies; novel therapeutics; novel vaccines; public health relevance; quantum

 DESCRIPTION (provided by applicant): Polychromatic flow cytometry (FC) is one of the most powerful analytical techniques routinely used by both basic research and clinical diagnostics laboratories for the immunological categorization of cells. Dyes used for FC typically exhibit broad fluorescent emission bands with full-width-at-half-maximum (fwhm) values of 50-80 nm. This limits the maximum number of dyes, and thus the number of cell biomarkers, that can be resolved in a given experiment. Compensating for spectral overlap between dyes is currently viewed as a necessary part of experimental design, requiring extensive pre-assay experimentation and mathematical compensation, thereby introducing experimental error and reducing sensitivity. Bacteriochlorins are a unique class of fluorescent dyes that offer a solution for accurate multiplexing with minimal compensation due to their very narrow emission bands (fwhm of 25-35 nm), typically less than half the spectral width of existing dyes. Through chemical modification, they can be tuned to emission wavelengths from the far red through the near-infrared (NIR) spectrum (700-900 nm). In addition, bacteriochlorins share a common excitation band, making possible the development of a full spectrum of NIR dyes excited by a single UV light source. To render bacteriochlorin dyes commercially viable, Phase II efforts will be focused on: 1) improving and expanding the bacteriochlorin dye portfolio and synthesis methods; 2) validating dye performance in FC panels with NIRvana Sciences' collaborators: a leading flow cytometry company and the Vaccine Research Center of the National Institute of Allergy and Infectious Diseases, and 3) developing procedures, methods, and protocols for commercial scale manufacturing. This Phase II STTR proposal is intended to continue the refinement and transfer of bacteriochlorin technology from North Carolina State University to NIRvana Sciences for commercial development, resulting in a bacteriochlorin dye portfolio with high impact potential for polychromatic FC. This enhanced multiplex capability will advance not only basic immunology research, but it will also accelerate novel vaccine and adjuvant discovery for HIV, malaria, tuberculosis, and emerging infectious disease threats. Greater multiplexing also is critical for analyzing reduced volume samples, for single cell studies, and for high throughput, high-resolution analyses.

 描述（由申请人提供）：多色流式细胞术（FC）是基础研究和临床诊断实验室常规用于细胞免疫学分类的最强大的分析技术之一。用于 FC 的染料通常表现出宽的荧光发射带，半峰全宽 (fwhm) 值为 50-80 nm。这限制了在给定实验中可以解析的染料的最大数量，从而限制了细胞生物标志物的数量。补偿染料之间的光谱重叠目前被视为实验设计的必要部分，需要大量的预测定实验和数学补偿，从而引入实验误差并降低灵敏度。菌绿素是一类独特的荧光染料，可提供解决方案 由于其发射带非常窄（半峰宽为 25-35 nm），通常小于现有染料光谱宽度的一半，因此可以以最小的补偿进行精确的多路复用。通过化学修饰，它们可以调整为从远红光到近红外 (NIR) 光谱（700-900 nm）的发射波长。此外，菌绿素具有共同的激发带，使得开发由单一紫外光源激发的全光谱近红外染料成为可能。为了使菌绿素染料商业化，第二阶段的工作将集中在：1）改进和扩展菌绿素染料组合和合成方法； 2) 与 NIRvana Sciences 的合作者验证 FC 面板中的染料性能：一家领先的流式细胞术公司和国家过敏和传染病研究所的疫苗研究中心，以及 3) 开发商业规模生产的程序、方法和方案。该第二阶段 STTR 提案旨在继续改进菌绿素技术，并将其从北卡罗来纳州立大学转移到 NIRvana Sciences 进行商业开发，从而形成对多色 FC 具有高影响潜力的菌绿素染料组合。这种增强的多重能力不仅将推进基础免疫学研究，还将加速针对艾滋病毒、疟疾、结核病和新出现的传染病威胁的新型疫苗和佐剂的发现。更大的多重性对于分析减少体积的样品、单细胞研究以及高通量、高分辨率分析也至关重要。