Development Of A Cytokine Chip

细胞因子具有多种调节功能，其中最重要的功能之一是调节T细胞分化。分泌的细胞因子谱已被证明是预测不同的T细胞分化，特别是调节细胞介导的和体液免疫的细胞因子。为了测量不同的细胞因子的个人样本中，同时，一个基本的细胞因子芯片已经开发的基础上固相免疫亲和提取的分析物的利益。重组链霉亲和素斑点的阵列通过羰基二咪唑桥衍生到硅烷化玻璃显微镜载玻片的表面。抗细胞因子抗体通过其碳水化合物部分被生物素化并与链霉亲和素结合。在分析之前用激光染料标记样品，并与芯片一起孵育。通过洗涤除去非反应性材料后，在扫描激光密度计中读取结合的分析物。发现芯片对所研究的所有五种细胞因子的灵敏度为100 - 150 pg/ml。进一步的研究需要优化芯片传感器和阅读机制，以进一步提高灵敏度。选择性研究表明，该芯片能够高度的特异性时，使用混合细胞因子样品。新的，增加的选择性芯片的发展打开了大门，研究一些重要的调控途径，如造血，炎症神经源性调节，伤口愈合同时在同一芯片上。未来的模型预计将包含30 - 100个配体，使多种分析成为可能。

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. Sensitivity of the chip was found to be 100 - 150 pg/ml for all five cytokines studied. Further studies require optimization of the chip sensor and reading mechanism to further increase sensitivity. Selectivity studies indicated that the chip was capable of a high degree of specificity when using mixed cytokine samples. Development of new, increased selectivity chips open the door to studying a number of important regulatory pathways such as hematopoesis, inflammation neurogenic regulation, and wound healing simultaneously on the same chip. Future models are expected to contain 30-100 ligands making multiple analyses possible.