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

: Neubauer, J.C.; Sébastien, I.; Germann, A.; Müller, S.C.; Meyerhans, A.; Briesen, H. von; Zimmermann, H.

Fulltext urn:nbn:de:0011-n-4382220 (1.7 MByte PDF)
MD5 Fingerprint: ab5377c7c52200fc1ebf9652ea86f7e6
Created on: 9.3.2017

Cytotechnology 69 (2017), No.1, pp.57-73
ISSN: 0920-9069
ISSN: 1573-0778
Journal Article, Electronic Publication
Fraunhofer IBMT ()

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.