Evaluation of a digital dispenser for direct curve dilutions in a vaccine potency assay.

Autor: Roselle C; Merck & Co. Inc., Kenilworth, NJ. Electronic address: christopher.roselle@merck.com., Whitehouse D; Merck & Co. Inc., Kenilworth, NJ. Electronic address: dana.whitehouse@merck.com., Follmer T; Merck & Co. Inc., Kenilworth, NJ. Electronic address: thy_follmer@merck.com., Ansbro F; Merck & Co. Inc., Kenilworth, NJ. Electronic address: frances_ansbro@merck.com., Bouaraphan S; Merck & Co. Inc., Kenilworth, NJ. Electronic address: Silikhone_bouaraphan@merck.com., Guan L; Merck & Co. Inc., Kenilworth, NJ. Electronic address: liming_guan@merck.com., Wang SK; Merck & Co. Inc., Kenilworth, NJ. Electronic address: shake_wang@merck.com., Shank-Retzlaff M; Merck & Co. Inc., Kenilworth, NJ. Electronic address: mary_retzlaff@merck.com., Verch T; Merck & Co. Inc., Kenilworth, NJ. Electronic address: thorsten_verch@merck.com.
Jazyk: angličtina
Zdroj: Journal of immunological methods [J Immunol Methods] 2017 Mar; Vol. 442, pp. 20-28. Date of Electronic Publication: 2016 Dec 27.
DOI: 10.1016/j.jim.2016.12.003
Abstrakt: Dilutions are a common source of analytical error, both in terms of accuracy and precision, and a common source of analyst mistakes. When serial dilutions are used, errors compound, even when employing laboratory automation. Direct point dilutions instead of serial dilutions can reduce error but is often impractical as they require either large diluent volumes or very small sample volumes when performed with traditional liquid handling equipment. We evaluated preparation of dilution curves using a picoliter digital dispenser, the HP, Inc. / TECAN D300 which is capable of accurately delivering picoliter volumes directly into sample wells filled with assay diluent. Dilution linearity and variability of the direct dilutions were similar to or less than those generated with a traditional liquid handler as measured using a fluorophore assay and an ELISA used to measure vaccine potency. Minimum concentrations for detergent in the dispensed sample were identified but no correlation with detergent characteristics was observed. The tolerance to protein in the sample was evaluated as well with up to 5% BSA having no impact on dispense linearity and precision. We found the digital dispenser to reduce automation complexity while maintaining or improving assay performance in addition to facilitating complex plate lay-outs.
(Copyright © 2017 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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