Evaluation of a digital dispenser for direct curve dilutions in a vaccine potency assay
Autor: | Christopher Roselle, Dana Whitehouse, Mary Shank-Retzlaff, Sha-Ke Wang, Thorsten Verch, Silikhone Bouaraphan, Liming Guan, Thy Follmer, Fran Ansbro |
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Rok vydání: | 2017 |
Předmět: |
0301 basic medicine
Accuracy and precision Serial dilution Sample (material) Detergents Immunology Enzyme-Linked Immunosorbent Assay 01 natural sciences Diluent 03 medical and health sciences Immunology and Allergy Vaccine Potency Fluorescent Dyes Automation Laboratory Miniaturization Chromatography Chemistry 010401 analytical chemistry Reproducibility of Results Small sample Equipment Design Reference Standards High-Throughput Screening Assays 0104 chemical sciences Dilution Spectrometry Fluorescence 030104 developmental biology Calibration Laboratory automation |
Zdroj: | Journal of Immunological Methods. 442:20-28 |
ISSN: | 0022-1759 |
Popis: | 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. |
Databáze: | OpenAIRE |
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