Development Of A Cytokine Chip

细胞因子芯片的开发

• 批准号: 7593817
• 负责人: Terry M. Phillips
• 金额: $ 1.54万
• 依托单位: NATIONAL INSTITUTE OF BIOMEDICAL IMAGING AND BIOENGINEERING
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7593817
• 关键词: Activated Lymphocyte; Antibodies; Binding; Carbohydrates; Cell Differentiation process; Cells; Cerebrospinal Fluid; Cerebrum; Characteristics; Chemistry; Childhood; Clinical; Commit; Critical Care; Development; Disease; Dyes; Evaluation; Excision; Exhibits; Glass; Growth Factor; Human; Humoral Immunities; Immobilization; Incubated; Individual; Infant; Inflammatory; Label; Lasers; Lung diseases; Lymphocyte; Measures; Mediating; Microscope; Modeling; Neurogenic Inflammation; Neuropeptides; Pain; Pathway interactions; Patients; Phase; Reader; Reading; Recombinants; Regulation; Regulatory Pathway; Sampling; Scanning; Slide; Solid; Specificity; Spottings; Staging; Streptavidin; Surface; System; T-Lymphocyte; Techniques; Undifferentiated; United States National Institutes of Health; Work; Wound Healing; base; cell growth regulation; clinical application; cytokine; design; immune function; improved; interest; Activated Lymphocyte; Antibodies; Binding; Carbohydrates; Cell Differentiation process; Cells; Cerebrospinal Fluid; Cerebrum; Characteristics; Chemistry; Childhood; Clinical; Commit; Critical Care; Development; Disease; Dyes; Evaluation; Excision; Exhibits; Glass; Growth Factor; Human; Humoral Immunities; Immobilization; Incubated; Individual; Infant; Inflammatory; Label; Lasers; Lung diseases; Lymphocyte; Measures; Mediating; Microscope; Modeling; Neurogenic Inflammation; Neuropeptides; Pain; Pathway interactions; Patients; Phase; Reader; Reading; Recombinants; Regulation; Regulatory Pathway; Sampling; Scanning; Slide; Solid; Specificity; Spottings; Staging; Streptavidin; Surface; System; T-Lymphocyte; Techniques; Undifferentiated; United States National Institutes of Health; Work; Wound Healing; base; cell growth regulation; clinical application; cytokine; design; immune function; improved; interest

Cytokines serve a variety of regulatory functions, one of the most important being the regulation of T-cell differentiation. Secreted cytokine profiles have been demonstrated to be predictive of different T-cell differentiation, especially cytokines regulating cell-mediated and humoral immunity. In order to measure different cytokine profiles simultaneously in individual samples, a basic cytokine chip has been developed based on solid-phase immunoaffinity extraction of the analytes of interest. Arrays of recombinant streptavidin spots were derivatized to the surfaces of silanized glass microscope slides via a carbonyldiimidazole bridge. Anti-cytokine antibodies were biotinylated via their carbohydrate moieties and bound to the steptavidin. Samples were labeled with a laser dye prior to analysis and incubated with the chip. Following removal of non-reactive materials by washing, the bound analytes were read in a scanning laser densitometer. Improvements in both the immobilization chemistry and techniques for keeping the antibody molecules active have increased the sensitivity of the chip to 1.0-2.5 pg/ml for all 50 cytokines studied. Studies are continuing to improve this level of sensitivity as well as further optimize the chip reading mechanism. The chip exhibits very good selectivity and a high degree of specificity when using both model mixtures of cytokines and human clinical samples. The chip is now in the evaluation stage prior to its use in such applications as the analysis of important regulatory pathways, including hematopoesis, inflammation neurogenic regulation, cellular regulation, and wound healing. Work has continued to develop and improve a static antibody array for measuring cytokines and growth factors in clinical samples. The acquisition of an automated chip reader greatly improved both sensitivity and selectivity of the antibody arrays. Additionally, the use of a dual laser system has expanded the usefulness of the array. Collaborative studies recently established with the Critical Care Center of the NIH Clinical Center have opened up new clinical applications for the antibody chip. At present, a panel of 50-100 specific antibodies is being immunochemically evaluated for inclusion in a patient-screening chip. This chip will be used to screen pediatric patients with inflammatory lung disease for regulatory cytokines, pain-associated neuropeptides, and other modulators associated with immune function in respiratory disease.

细胞因子具有多种调节功能，最重要的是T细胞分化的调节。 已证明分泌的细胞因子谱图可以预测不同的T细胞分化，尤其是调节细胞介导的和体液免疫的细胞因子。 为了同时测量各个样品中的不同细胞因子谱，基于感兴趣的分析物的固相免疫疗法提取，已经开发了基本的细胞因子芯片。 重组链霉亲和素斑点的阵列通过羰基二咪唑桥衍生在硅烷化玻璃显微镜载玻片的表面上。 抗周期因子抗体通过其碳水化合物部分进行生物素化，并与steptavidin结合。 在分析之前，将样品用激光染料标记，并与芯片孵育。 通过洗涤去除非反应性材料后，在扫描激光密度计中读取结合的分析物。 固定化学和保持活性抗体分子的技术的改善都使芯片的敏感性提高到了所有研究的50个细胞因子的敏感性至1.0-2.5 pg/ml。 研究正在继续提高这种敏感性，并进一步优化芯片阅读机制。 当使用细胞因子和人类临床样品的模型混合物时，芯片具有很好的选择性和高度特异性。 现在，芯片在使用之前在评估阶段处于评估阶段，例如对重要调节途径的分析，包括血液素，炎症神经源性调节，细胞调节和伤口愈合。 工作继续开发和改善用于测量临床样品中细胞因子和生长因子的静态抗体阵列。 自动芯片读取器的获取极大地提高了抗体阵列的灵敏度和选择性。 此外，使用双激光系统扩大了数组的实用性。 NIH临床中心的重症监护中心最近建立的合作研究为抗体芯片开辟了新的临床应用。 目前，正在对50-100个特异性抗体进行免疫化学评估，以纳入患者筛查芯片中。 该CHIP将用于筛查儿科患者患有炎症性肺部疾病，以进行调节性细胞因子，疼痛相关的神经肽以及与免疫功能相关的其他调节剂在呼吸道疾病中。