Simultaneous analysis of cell-borne and soluble biomarkers by high throughput acoustic flow cytometry

项目摘要 分析复杂的细胞过程，以了解疾病，药物开发或测试 治疗的有效性通常需要动态测量细胞标志物和细胞内的 产品.在复杂的细胞中同步和快速进行这种测量的能力- 包含使用单一仪器分析的样品将是有用和有益的 生物技术.该项目的目标是使用免疫组织化学的方法， 免疫分型和定量监测其分泌的细胞因子作为重要模型 系统悬浮细胞样品的高通量测量是可能的， BennuBio的Velocyt®多流声学流式细胞术系统。这个项目的主要创新之处在于， 该项目是实施负声学对比颗粒（NACPs）作为一个基于珠 免疫分析平台，显著扩展了Velocyt®仪器的分析能力 用于同时分析细胞相关和可溶性生物标志物。与细胞相反， NACP迁移到Velocyt®流动流中的离散位置并在这些位置进行分析， 分离到流动流中的不同位置，允许容易地区分珠粒和 单个样本中的细胞。在这个概念验证阶段I研究中，我们的目标是同时进行 在相同条件下测量CD 4和CD 8群体中的T细胞活化标志物CD 25， 我们测量分泌的干扰素-γ和肿瘤坏死因子-α的时间。虽然这次示威将 对开发用于治疗一系列免疫性疾病的药物具有重要意义 以及涉及T细胞活化的传染病，它也将提供一个坚实的基础， 设计和商业化可定制的基于NACP的试剂盒，以解决广泛的 跨生物技术学科。

Project Summary Analysis of complex cellular processes in understanding disease, development of drugs, or testing efficacy of therapies often requires dynamic measurements of both cellular markers and cellular products. The ability to synchronously and rapidly conduct such measurements in complex cell- containing samples using a single instrumental analysis will be useful and beneficial across biotechnology. The goal of this project is to enable such multiplex analysis of T-cell activation using immunophenotyping and quantitative monitoring of their secreted cytokines as an important model system. High throughput measurement of samples of suspended cells is made possible using BennuBio’s Velocyt® multi-stream acoustic flow cytometry system. The primary innovation of this project is the implementation of negative acoustic contrast particles (NACPs) as a bead-based immunoassay platform that significantly extends the analytical capabilities of the Velocyt® instrument for simultaneous analysis of cell-associated and soluble biomarkers. In contrast to cells, which migrate to, and are analyzed at, discreet positions within the Velocyt® flow stream,NACPs are separated to distinct positions in the flow stream, allowing for easy discrimination between bead and cells in a single sample. In this proof-of-concept Phase I research, we aim to conduct simultaneous measurement of the T-cell activation marker CD25 in both the CD4 and CD8 population at the same time that we measure secreted interferon-γ and tumor necrosis factor-α . While this demonstration will be of significance to the development of pharmaceuticals for treatment of a range of immunological and infectious diseases that involve T-cell activation, it will also provide a firm basis by which to design and commercialize customizable NACP-based reagent kits to address a wide range of disciplines across biotechnology.