在过去的几年里，科学界已经在推进我们对免疫系统的理解方面取得了巨大进展，从其各种组成部分到这些组件内运营的分子途径的基本功能。凭借新的，最先进的工具和技术，免疫医生现在有能力更好地了解对各种抗原的免疫应答机制，从而使他们在进行新的治疗免疫系统相关疾病的方法中的发展中的发展设计疫苗，用于打击传染病和癌症。
目前，免疫细胞检测，测量和功能分析的最敏感的技术之一是酶联免疫蛋白（ELISPOT.测定。Covanceuses the ELISPOT technique in applications such as evaluation of vaccine efficacy and immunogenicity of biological products.
Benefits of ELISPOT assays
The ELISPOT assay is one of the most sensitive in vitro cellular assays for detecting and enumerating individual immune cells that secrete soluble mediators in response to a stimulus. The ELISPOT assay was originally developed for analyzing specific antibody-secreting cells based on theenzyme-linked immunosorbent assay (ELISA)but it has been adapted to measure the frequencies of cells that produce and secrete other effector molecules such as cytokines.
取决于分析的可溶因子，ELISPOT测定比常规ELISA测定更敏感在200至400倍之间。它能够检测到300,000个低频细胞因子分泌细胞中最多1个。由于响应于抗原释放的细胞因子可以映射到单个电池，因此可以使用该技术确定T细胞响应频率。ELISPOT测定还提供了一种被引发的细胞因子反应类型的指示，因此是免疫应答的类型。与细胞内细胞因子染色如诸如细胞内细胞因子染色的其他方法相比，ELISPOT测定可以产生更好的定量和定性数据。
Applicability Across Multiple Immunology Disciplines
ELISPOT.testing can be used across several disciplines of immunology, and is particularly useful in areas where the ex vivo measurement of low-frequency T cell responses is required. For instance, ELISPOT assays are useful in monitoring the frequency and profile of circulating donor-reactive T cells in organ transplantation; measuring cytotoxic T lymphocyte (CTL) activity in cancer research; measuring memory responses in infectious diseases and vaccine development; and detecting autoreactive cells (which typically occur at low frequencies) in autoimmune disease.
诸如广泛用于治疗包括癌症和自身免疫的各种疾病的生物治疗剂是能够诱导不需要的免疫反应。这种不需要的免疫应答可以以激活的免疫细胞的形式表现出可导致非常高水平的炎性细胞因子的分泌，这一现象称为“细胞因子风暴”。'细胞因子风暴'可以产生严重后果，包括死亡。ELISPOT测定也可用于评估生物学是否能够在其在动物和人类使用之前引发“细胞因子风暴”。
维护ELISPOT绩效标准
The multiple, complex steps of an ELISPOT assay require highly specialized capabilities. The following are some factors that can impact ELISPOT assay performance.
Cell recovery and viability:标准化ELISPOT测定中的最关键步骤之一是确保从冷冻保存后拯救外周血单核细胞（PBMC）后的充足的细胞回收和活力。凭借其全球业务，Covance的中央实验室服务可以从测试对象和PBMCS将高质量的PBMC迅速准备和冷冻储存高质量的PBMC，以便在单一的测试位置，确保在ELISPOT等功能测定中的最佳性能。
Operator Technique and Proficiency:Carrying out ELISPOT assays can be laborious and can yield variable results. Covance has experienced scientists and associates trained specifically in ELISPOT technique, who perform these assays routinely using optimized and standardized protocols with minimal assay variability.
Result variability:In order to minimize variability in results and improve the ability to confidently compare subjects from different clinical trial sites, steps should be taken to ensure initial assay performance and adherence to clinical standards over time. In addition to following a standard operating procedure (SOP) for the execution of ELISPOT assays, Covance experts follow a documented training protocol, under a set of guiding ‘quality principles’. Our assay validation parameters ensure optimal assay performance and high quality data generation.
ELISPOT专业知识与全球范围有音乐会，是您选择的CRO合作伙伴的重要能力。有关更多信息，请访问Covance.com.
