RBC typing antibodies labeled with rare earth phosphor upconverting nanocrystals

稀土磷上转换纳米晶体标记的红细胞分型抗体

• 批准号: 7544346
• 负责人: Ajith Kumar
• 金额: $ 22.98万
• 依托单位: SUNSTONE BIOSCIENCES , INC.
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-01-26 至 2010-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7544346
• 关键词: Address; Adverse effects; Allergic Reaction; Antibodies; Antigens; Binding; Biological Assay; Blood; Blood Transfusion; Blood typing procedure; Businesses; Characteristics; Communities; Detection; Development; Diagnostics Research; Erythrocytes; Funding; Goals; Hemagglutination; Image; Incidence; Label; Life; Light; Manuals; Medical; Methods; Mission; Molecular; Monoclonal Antibodies; National Institute of Biomedical Imaging and Bioengineering; Optics; Patients; Pennsylvania; Performance; Phase; Phenotype; Photobleaching; Procedures; Property; Quantum Dots; Reaction; Reading; Reagent; Research; Resolution; Risk; Running; Sampling; Signal Transduction; Small Business Technology Transfer Research; Specific qualifier value; Specificity; System; Technology; Testing; Time; Transfusion; United States National Institutes of Health; Universities; Work; antibody conjugate; base; design and construction; fluorophore; imaging modality; meetings; monitoring device; nanocrystal; nanoparticle; novel; physical property; programs; prototype; public health relevance; rapid detection; tool

DESCRIPTION (provided by applicant): The product that will result from the proposed STTR application from Sunstone Biosciences and the University of Pennsylvania is a novel labeled antibody-based reagent kit that can be used to rapidly identify diverse red blood cell phenotypes prior to blood transfusion. At present, pre-transfusion blood typing involves only the major antigens of the ABO and Rh groups, and relies on the use of a largely- manual hemagglutination reaction. The time- and labor-intensiveness of this procedure limits the number of RBC sub-groups that can be tested prior to a transfusion, risking the development of sensitization of patients who receive multiple transfusions throughout their life. We plan to develop a large panel of RBC antigen-specific MAbs, labeled with rare-earth nanophosphors, able to rapidly determine RBC phenotype using a wide range of common and rare antigens in an automatable, multiplexed assay format. The programmable, and narrow optical spectra of rare-earth crystals are ideal for generating molecular detection systems that are capable of vastly greater multiplexing than conventional optical probes. At the end of Phase 2 of the proposed program, we anticipate having designed and constructed a panel of approximately 20 or more antibodies, each with a unique optical emission signal, that can be used to create a revolutionary blood phenotyping "signature". The upconverting characteristics of the nanocrystals are unique identifiers of target-specific labeling and quantification. Other methods of molecular detection, including Quantum Dots and conventional fluorophores, are utilized widely throughout the research community but possess many limitations such as photobleaching, poor resolution, and the limited number of samples permitted to run simultaneously. Rare-earth phosphor nanocyrstals address these problems, as inert, stable light emitters that allow for high resolution and vastly multiplexed imaging. PUBLIC HEALTH RELEVANCE: Sunstones Biosciences has, with its consortium partners at Princeton University, generated preliminary evidence that nanocrystals as small as ~10 nm can be synthesized, that these nanocrystals retain their unique optical properties, and that they can be coated for functionalization (such as for conjugation to antibodies). In this Phase 1 proposal, we will extend these studies to generate and test at least three phosphor nanocrystal-labeled antibodies specific for RBC antigens. This proposal will reduce the incidence of allergic reactions and other side-effects of blood transfusion by developing a new product for comprehensively matching donor and recipient blood types. Unlike conventional methods for blood typing, the proposed method will allow rapid detection of even rare blood markers.

描述（由申请人提供）：Sunstone Biosciences和宾夕法尼亚大学提出的STTR申请将产生的产品是一种新型标记抗体试剂盒，可用于在输血前快速鉴定不同的红细胞表型。目前，输血前血型鉴定仅涉及ABO和Rh血型的主要抗原，并且依赖于大部分人工血凝反应的使用。该程序的时间和劳动密集性限制了输血前可以检测的RBC亚组的数量，从而有可能使一生中接受多次输血的患者发生致敏反应。我们计划开发一个大面板的RBC抗原特异性单克隆抗体，用稀土纳米荧光粉标记，能够快速确定RBC表型使用广泛的常见和罕见的抗原在自动化，多路复用的测定格式。稀土晶体的可编程和窄光谱是产生分子检测系统的理想选择，该系统能够比传统的光学探针进行更大的多路复用。在拟议计划的第二阶段结束时，我们预计已经设计并构建了一组约20种或更多种抗体，每种抗体都具有独特的光发射信号，可用于创建革命性的血液表型“签名”。纳米晶体的上转换特性是靶特异性标记和定量的唯一标识符。其他分子检测方法，包括量子点和传统荧光团，在整个研究界被广泛使用，但具有许多局限性，如光漂白，分辨率差，以及允许同时运行的样品数量有限。稀土磷光体纳米晶体解决了这些问题，作为惰性，稳定的光发射器，允许高分辨率和大量多路复用成像。公共卫生相关性：Sunstones Biosciences与其在普林斯顿大学的联盟合作伙伴一起产生了初步证据，表明可以合成小至10 nm的纳米晶体，这些纳米晶体保留了其独特的光学性质，并且它们可以被涂覆用于功能化（例如用于与抗体缀合）。在这个第一阶段的提案中，我们将扩展这些研究，以产生和测试至少三种对RBC抗原特异性的荧光纳米标记抗体。该提案将通过开发一种新产品来全面匹配供体和受体血型，从而减少过敏反应和其他输血副作用的发生率。与传统的血型分型方法不同，该方法将允许快速检测甚至罕见的血液标记物。