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

通过高通量声学流式细胞术同时分析细胞源性和可溶性生物标志物

基本信息

批准号：
10602869
负责人：
John Peter O'Rourke
金额：
$ 27.52万
依托单位：
BENNUBIO INC
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-03-01 至 2024-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10602869
关键词：
Acoustics Address Antibodies Autoimmune Diseases Biological Biological Assay Biological Markers Biological Models Biomedical Research Biotechnology Biotin Blood Blood Cells CD28 gene CD3 Antigens CD8-Positive T-Lymphocytes CD8B1 gene Cell model Cell physiology Cell secretion Cell surface Cells Clinical Communicable Diseases Complex Coupled Cytometry Data Detection Development Diagnosis Discipline Discrimination Disease Elastomers Engineering Exhibits Flow Cytometry Fluorescence Frequencies Goals Human IL2RA gene Immobilization Immune System Diseases Immunoassay Immunophenotyping Interferon Type II Interferons Lasers Liquid substance Measurement Measures Methodology Methods Microscopic Microspheres Modeling Monitor Oncology Pathway interactions Performance Peripheral Blood Mononuclear Cell Pharmacologic Substance Phase Plasma Population Positioning Attribute Preparation Process Prognosis Property Protein Analysis Proteins Reagent Research Sampling Serum Silicones Specificity Speed Stream Streptavidin Supporting Cell Surface Suspensions System T-Cell Activation T-Lymphocyte TNF gene Technology Testing Time Travel Treatment Efficacy Up-Regulation Variant Width Work Simplification aqueous commercialization cytokine design detection method diagnostic value drug development drug discovery efficacy testing elastomeric electric impedance innovation insight instrument migration multiplex assay novel particle pressure protein biomarkers receptor solute specific biomarkers stem cell technology therapeutic development therapeutic target tissue culture

项目摘要

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.

项目概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功效通常需要动态测量细胞标记物和细胞产品能够在复杂的细胞中同步快速地进行此类测量。使用单一仪器分析包含样品将是有用且有益的该项目的目标是利用生物技术对 T 细胞激活进行多重分析。免疫表型和分泌细胞因子的定量监测作为重要模型使用系统可以对悬浮细胞样品进行高通量测量。 BennuBio 的 Velocyt® 多流声学流式细胞术系统是该系统的主要创新。该项目是实施负声学对比粒子（NACP）作为基于珠子的免疫分析平台显着扩展了 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。迁移到 Velocyt® 流中的离散位置并进行分析，NACP 是分离到流中的不同位置，可以轻松区分珠子和在这一概念验证阶段的研究中，我们的目标是同时进行单个样本中的细胞。同时测量 CD4 和 CD8 群体中的 T 细胞激活标记 CD25 我们测量分泌的干扰素-γ 和肿瘤坏死因子-α 的时间，而本演示将进行。对于开发治疗一系列免疫性疾病的药物具有重要意义和涉及 T 细胞激活的传染病，它也将为设计和商业化基于 NACP 的可定制试剂盒，以满足广泛的需求跨生物技术学科。