Zellsortiersystem

细胞分选系统

• 批准号: 431349925
• 金额: --
• 依托单位: Universität zu Köln
• 依托单位国家: 德国
• 项目类别: Major Research Instrumentation
• 财政年份: 2019
• 资助国家: 德国
• 起止时间: 2018-12-31 至 无数据
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/431349925?language=en
• 关键词: Zellsortiersystem

Our work focuses on the analysis of B cell and antibody responses against clinically relevant pathogens such as HIV, HCV, or Ebola. We use our findings to develop antibody-based therapies and inform on novel vaccination strategies. In the last years, we have been able to identify highly potent antibodies that we also successfully tested in clinical trials. Our advanced single cell cloning methods critically rely on multicolor flow cytometry with bulk and single cell sorting applications. With these methods, we are able to isolate specific cell populations, such as naive or memory B cells, by labeling them with fluorescent dyes. Several labels are necessary to yield a pure B cell sub-population, which in turn requires an instrument with several excitation lasers, emission filters, and detectors. Since we work with infectious materials (BSL2), the instrument further needs to be operated within a safety cabinet. We already have all necessary permissions to conduct these kind of experiments, but are currently working on a rental instrument with a fixed expiration of the rental contract. Thus, we apply for funding for a flow cytometer with single cell sorting function in order to continue our intensive research on novel vaccination- and therapy strategies.We have optimized our single cell cloning methods in the past and currently reached a capacity limit due to technical and structural limitations, which hampers the realization of our ongoing and planed projects. Based on our experience in manual single cell cloning, we plan to build an automated single cell platform for antibody analysis and screening. With this, we aim to (i) increase the sample throughput, (ii) reduce overall costs by volume reduction, and (iii) ensure a high quality among all analyses independent of any experimenter. To this end, we apply for funding for a liquid handling robot, that is able to process several sample in a 384-well plate format in parallel. The automation will allow us to identify therapeutically applicable antibodies against established and emerging infectious diseases in a shorter time frame and thereby strengthen our national and international position as an antibody analysis center of excellence.

我们的工作重点是分析B细胞和抗体对临床相关病原体（如HIV、HCV或埃博拉病毒）的反应。我们利用我们的研究结果开发基于抗体的疗法，并为新型疫苗接种策略提供信息。在过去的几年里，我们已经能够识别出高效的抗体，我们也在临床试验中成功测试。我们先进的单细胞克隆方法主要依赖于流式细胞仪的批量和单细胞分选应用。利用这些方法，我们能够通过用荧光染料标记它们来分离特定的细胞群，例如幼稚或记忆B细胞。需要几种标记来产生纯的B细胞亚群，这又需要具有几种激发激光器、发射滤光器和检测器的仪器。由于我们使用感染性材料（BSL 2），因此仪器还需要在安全柜内操作。我们已经获得了进行此类实验的所有必要许可，但目前正在研究租赁合同到期的租赁仪器。因此，我们申请了一台具有单细胞分选功能的流式细胞仪的资助，以继续我们对新型疫苗接种和治疗策略的深入研究。我们在过去优化了我们的单细胞克隆方法，目前由于技术和结构的限制，达到了能力极限，这阻碍了我们正在进行和计划中的项目的实现。基于我们在人工单细胞克隆方面的经验，我们计划建立一个用于抗体分析和筛选的自动化单细胞平台。因此，我们的目标是（i）增加样品通量，（ii）通过减少体积来降低总体成本，以及（iii）确保所有分析的高质量，而不依赖于任何实验者。为此，我们申请资金用于液体处理机器人，该机器人能够并行处理384孔板格式中的多个样品。自动化将使我们能够在更短的时间内识别针对已建立和新出现的传染病的治疗适用抗体，从而加强我们作为抗体分析卓越中心的国家和国际地位。