A Celigo Whole Well Imaging Cytometer for the Crick Covid neutralisation pipeline

用于 Crick Covid 中和管道的 Celigo Whole Well 成像细胞仪

• 批准号: MR/X012883/1
• 负责人: Michael Howell
• 金额: $ 25.63万
• 依托单位: The Francis Crick Institute
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2022
• 资助国家: 英国
• 起止时间: 2022 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FX012883%2F1
• 关键词: Celigo; Whole; Well; Imaging; Cytometer

The Crick achieves operational and research efficiencies and economies-of-scale through centralised facilities and functions, known as Science Technology Platforms, that provide all researchers at the Crick, irrespective of affiliation, with access to cutting-edge equipment for research and laboratory enabling functions such as the High Throughput Screening facility (HTS).The Crick Covid Surveillance unit (CCSU) is an offshoot of the HTS facility and was established to run a pipeline of activity delivering a high throughput live-virus antibody neutralisation assay to measure the ability of patient serum to prevent cell infection by SARS-CoV-2 across thousands of samples simultaneously. This unique neutralisation assay platform (developed in the HTS facility) is essential to many ongoing research studies, for example those assessing the likely impact of Variants of Concern on current vaccinated populations and in clinically vulnerable cohorts. The pipeline is able to generate neutralization data against new VOCs (e.g. Delta and Omicron) within 2-3 weeks upon receipt of a novel virus sample and can process thousands of samples per week at GCP level. Data from this pipeline has measured neutralising antibody titres against Delta and Omicron variants in participants in the Crick-Legacy study (recipients of both the Pfizer and Oxford-AstraZeneca vaccines). These data are shared with UK government scientific advisory boards, contributing to the extension of UK pandemic restrictions to allow more people to receive a second vaccine dose in summer 2021 and the successful booster campaign. Publications reporting these findings have been cited nearly 600 times. This pipeline also enabled the comparison of antibody titres in healthy adults with those of both cancer patients (Crick-CAPTURE study) and kidney dialysis patients (Crick-NAOMI study), providing important data on prioritisation of vulnerable patient groups. In addition, data on the in vitro neutralising efficacy of the synthetic monoclonal antibody, Sotrovimab was shared with NHSE and the Chief Medical Officer and supported the ongoing use of this drug for patients infected with Omicron BA.1 and BA.2. There are no other similar pipelines capable of this scale of live virus assay in the UK (possibly internationally) and with near real time reporting, the data produced remains a vital tool in the UK and international pandemic preparedness and response.Currently, the neutralisation pipeline is made robust by using multiple HTS liquid handling devices to ensure continuity of delivery. However, the primary imaging (and measurements derived from them) is restricted to one microscope, the Opera Phenix. This reliance on one imaging device is a weak point for this pipeline. This proposal is to deploy the Celigo scanning cytometer in this assay and thereby remove the dependence on one machine for a pipeline that needs to remain operational. Moreover, acquisition on the Celigo is 4-5 times faster meaning that pipeline throughput and capacity can be significantly increased.The Covid neutralisation pipeline accounted for 50% of HTS usage over 2020/21. Having a high-end microscope occupied with one type of assay necessarily restricts access by other users (currently measured in weeks of delay for access to the Opera Phenix). By moving the primary imaging platform for the CCSU activity to the Celigo, we would greatly increase the availability of high content microscopy to other Crick researchers and thereby improve service delivery for more than 100 Crick researchers who used the HTS facility over 20/21. The availability of a Celigo imager will also enable improvements to existing activities requiring analysis of cell populations (at scale) in the areas of radiation biology, stem cell biology and general cell viability. In addition, there will be new opportunities for users to explore automation of previously manual assays e.g. Influenza plaque assays

克里克通过集中的设施和功能实现了运营和研究效率以及规模经济，这些设施和功能被称为科学技术平台，为克里克的所有研究人员，无论隶属关系如何，能够使用尖端设备进行研究和实验室支持功能，例如高通量筛查设施（HTS）。克里克新冠病毒监测单位（CCSU）是HTS设施的一个分支，其建立是为了运行提供高通量活病毒抗体中和测定的活动管道，以同时测量患者血清在数千个样品中预防SARS-CoV-2细胞感染的能力。这种独特的中和试验平台（在HTS设施中开发）对于许多正在进行的研究至关重要，例如评估关注变体对当前接种疫苗人群和临床易感人群的可能影响的研究。该管道能够在收到新病毒样本后2-3周内生成针对新VOC（例如Delta和Omicron）的中和数据，并且可以在GCP水平上每周处理数千个样本。来自该管道的数据测量了Crick-Legacy研究参与者（辉瑞和牛津-阿斯利康疫苗接种者）针对Delta和Omicron变体的中和抗体滴度。这些数据与英国政府科学咨询委员会共享，有助于延长英国大流行限制，使更多人能够在2021年夏季接种第二剂疫苗，并成功开展加强疫苗接种活动。报道这些发现的出版物被引用了近600次。该管道还可以将健康成人的抗体滴度与癌症患者（Crick-CAPTURE研究）和肾透析患者（Crick-NAOMI研究）的抗体滴度进行比较，为弱势患者群体的优先级提供重要数据。此外，与NHSE和首席医学官共享了合成单克隆抗体Sotrovimab的体外中和效力数据，并支持该药物持续用于Omicron BA.1和BA.2感染患者。在英国（可能是国际上）没有其他类似的管道能够进行这种规模的活病毒检测，并且由于接近真实的时间报告，所产生的数据仍然是英国和国际大流行病准备和应对的重要工具。目前，通过使用多个HTS液体处理设备来确保输送的连续性，中和管道变得强大。然而，主要成像（以及由此产生的测量）仅限于一台显微镜，即Opera Phenix。这种对一个成像设备的依赖是该流水线的弱点。该提案旨在将Celigo扫描细胞仪部署在该分析中，从而消除对需要保持运行的管道的一台机器的依赖。此外，收购Celigo的速度快了4-5倍，意味着管道吞吐量和容量可以显著增加。2020/21年度，新冠病毒中和管道占HTS使用量的50%。高端显微镜被一种类型的分析占用必然会限制其他用户的访问（目前以访问Opera Phenix的延迟数周来衡量）。通过将CCSU活动的主要成像平台转移到Celigo，我们将大大增加其他Crick研究人员的高内容显微镜的可用性，从而改善20/21使用HTS设施的100多名Crick研究人员的服务交付。Celigo成像仪的可用性还将有助于改进需要在辐射生物学、干细胞生物学和一般细胞活力领域分析细胞群（规模）的现有活动。此外，用户将有新的机会探索以前手动检测的自动化，例如流感菌斑检测