Towards standardized automated immunomonitoring: an automated ELISpot assay for safe and parallelized functionality analysis of immune cells

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• 作者：J. C. Neubauer ; I. Sébastien ; A. Germann ; S. C. Müller ; A. Meyerhans…
• 关键词：ELISpot ; Automation ; Liquid classes ; Operator safety ; HIV vaccines ; PBMC
• 刊名：Cytotechnology
• 出版年：2017
• 出版时间：February 2017
• 年：2017
• 卷：69
• 期：1
• 页码：57-73
• 全文大小：1941KB
• 刊物类别：Chemistry and Materials Science
• 刊物主题：Biotechnology; Biomedicine, general; Biochemistry, general;
• 出版者：Springer Netherlands
• ISSN：1573-0778
• 卷排序：69

文摘

The ELISpot assay is used for the detection of T cell responses in clinical trials and vaccine evaluations. Standardization and reproducibility are necessary to compare the results worldwide, inter- and intra-assay variability being critical factors. To assure operator safety as well as high-quality experiment performance, the ELISpot assay was implemented on an automated liquid handling platform, a Tecan Freedom EVO. After validation of the liquid handling, automated loading of plates with cells and reagents was investigated. With step by step implementation of the manual procedure and liquid dispensing optimization on the robot platform, a fully automated ELISpot assay was accomplished with plates remaining in the system from the plate blocking step to spot development. The mean delta difference amounted to a maximum of 6%, and the mean dispersion was smaller than in the manual assay. Taken together, we achieved with this system not only a lower personnel attendance but also higher throughput and a more precise and parallelized analysis. This platform has the potential to guarantee validated, safe, fast, reproducible and cost-efficient immunological and toxicological assays in the future.