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

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: 8718340
• 负责人: James Bruce Pitner
• 金额: $ 22.5万
• 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 2015-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8718340
• 关键词: Antibodies; Area; Autoimmunity; Basic Science; Binding; Biological Assay; Bronchoscopy; Buffers; CD4 Positive T Lymphocytes; CD8B1 gene; Cell Count; Cerebrospinal Fluid; Childhood; Clinical; Color; Communicable Diseases; Companions; Complex; Core Biopsy; Core Facility; Diagnosis; Diagnostic; Dyes; Endoscopy; Esters; Exhibits; Financial compensation; Fine needle aspiration biopsy; Flow Cytometry; Fluorescence; Fluorescent Dyes; Foundations; HIV Seropositivity; Hematologic Neoplasms; Hematological Disease; Hematopoietic Neoplasms; Human; Immunology; Immunophenotyping; In Vitro; Incidence; Intellectual Property; Laparoscopy; Lasers; Legal patent; Light; Lymphoma; Measurement; Methodology; NIH Vaccine Research Center; Nature; Pathway interactions; Patients; Play; Positioning Attribute; Procedures; Reliance; Role; Route; Running; Sample Size; Sampling; Science; Scientist; Solutions; Specimen; Speed; Staining method; Stains; System; T-Lymphocyte; T-Lymphocyte Subsets; Techniques; Technology; Temperature; Testing; Titrations; Transplantation; Water; Width; Work; absorption; bacteriochlorin; base; cancer diagnosis; cellular imaging; clinical application; clinical assay development; commercialization; cost; design; disease diagnosis; fluorophore; in vivo; indexing; innovation; leukemia; leukemia/lymphoma; member; minimally invasive; public health relevance; research study; transplantation typing; water solubility

DESCRIPTION (provided by applicant): Polychromatic flow cytometry [FC] is one of the most powerful analytical techniques routinely used in immunology. Flow cytometry plays a critical role in several growing clinical applications: leukemia and lymphoma immunophenotyping for diagnosis and sub-categorization (now using 8-10 colors); in CD4/CD8 measurement in HIV-positive patients; and in HLA typing for transplantation cross-matching. With the increasing incidence of hematologic malignancies and use of transplantation, the number of FC tests will continue to grow as reliance on the methodology deepens. Increasing the number of fluorophores on a single sample will give clinicians a higher level of accuracy in the diagnosis and management of certain pediatric hematological disorders, where sample size is limited, and even allow for sub-categorization of the leukemia or lymphoma. Further utility of additional fluorophores for these polychromatic panels is for diagnostic samples of small volume such as specimens from an ever-increasing number of minimally invasive procedures that produce limited volume specimens, such as fine needle aspiration, laparoscopy, core biopsy, endoscopy, bronchoscopy, and notoriously hypocellular cerebrospinal fluid. Due to the small sample size of most diagnostic specimens in current practice, often insufficient cell numbers are available for the necessarily numerous panels to reach a diagnosis. A uniform deficiency of most common fluorophores is their broad fluorescence [emission bands], which cause spectral overlap, requiring extensive pre-assay experimentation and use of mathematical compensation, all of which increase experimental error, reduce sensitivity, and limit multiplexing. NIRvana Sciences, Inc. proposes to develop the first three members of a palette of water-soluble, conjugatable, bright, emission wavelength-tunable bacteriochlorin fluorophores characterized by sharp emission bands in the near-infrared [NIR] region upon excitation by commonly used laser wavelengths. The new fluorophores are based on a platform technology enabled by concise, versatile synthetic routes to bacteriochlorins with the following features: photo and thermal stability; rational control of excitation and emission band positions; additional substitution for water solubility and bioconjugation; and light absorption at laser lines of 355, 532, and 561 nm [with corresponding intense NIR emission bands]. The bacteriochlorins exhibit long "Stokes'-like shifts" as all three absorption bands produce the same strong emission in the NIR region. These new fluorescent dyes fill an important unmet need in cancer diagnosis as well as pediatric and other limited-sample applications.

描述（由申请方提供）：多色流式细胞术[FC]是免疫学中常规使用的最强大的分析技术之一。流式细胞术在几个日益增长的临床应用中发挥着关键作用：白血病和淋巴瘤免疫表型诊断和亚分类（现在使用8-10种颜色）; HIV阳性患者的CD 4/CD 8测量;以及移植交叉配型的HLA分型。随着恶性血液病发病率的增加和移植的使用，随着对方法学依赖的加深，FC检测的数量将继续增长。增加单个样本上的荧光团数量将使临床医生在样本量有限的某些儿科血液疾病的诊断和管理中具有更高的准确性，甚至允许对白血病或淋巴瘤进行亚分类。用于这些多色面板的附加荧光团的进一步用途是用于小体积的诊断样品，例如来自不断增加的产生有限体积样本的微创程序的样本，例如细针抽吸、腹腔镜检查、芯活检、内窥镜检查、支气管镜检查和众所周知的细胞过少的脑脊液。由于目前实践中大多数诊断标本的样本量较小，因此通常没有足够的细胞数量可用于必要的众多面板以达到诊断。大多数常见荧光团的一个统一缺陷是它们的宽荧光[发射带]，这会导致光谱重叠，需要大量的预测定实验和使用数学补偿，所有这些都会增加实验误差，降低灵敏度并限制多路复用。NIRCRAFT Sciences，Inc.提出开发水溶性的、可缀合的、明亮的、发射波长可调谐的菌绿素荧光团的调色板的前三个成员，其特征在于在被常用的激光波长激发时在近红外[NIR]区域中具有尖锐的发射带。新的荧光团是基于一个平台技术，通过简洁，通用的合成路线，以细菌绿素具有以下特点：光和热稳定性;合理控制激发和发射带的位置;额外的替代水溶性和生物共轭;和光吸收在355，532和561 nm的激光线[与相应的强近红外发射带]。细菌绿素表现出长的“斯托克斯样位移”，因为所有三个吸收带在NIR区域产生相同的强发射。这些新的荧光染料填补了癌症诊断以及儿科和其他有限样本应用中重要的未满足需求。

项目成果

Bioconjugatable, PEGylated Hydroporphyrins for Photochemistry and Photomedicine. Narrow-Band, Near-Infrared-Emitting Bacteriochlorins.

用于光化学和光医学的生物共轭聚乙二醇化氢卟啉。

• DOI: 10.1039/c6nj01155a
• 发表时间: 2016
• 期刊: New journal of chemistry = Nouveau journal de chimie
• 作者: Zhang,Nuonuo;Jiang,Jianbing;Liu,Mengran;Taniguchi,Masahiko;Mandal,AmitKumar;Evans-Storms,RosemaryB;Pitner,JBruce;Bocian,DavidF;Holten,Dewey;Lindsey,JonathanS
• 通讯作者: Lindsey,JonathanS