Evaluation of a digital dispenser for direct curve dilutions in a vaccine potency assay
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- 作者：Christopher Roselle christopher.roselle@merck.com ; Dana Whitehouse dana.whitehouse@merck.com ; Thy Follmer thy_follmer@merck.com ; Fran Ansbro frances_ansbro@merck.com ; Silikhone Bouaraphan Silikhone_bouaraphan@merck.com ; Liming Guan liming_guan@merck.com ; Sha-Ke Wang shake_wang@merck.com ; Mary Shank-Retzlaff mary_retzlaff@merck.com ; Thorsten Verch ; thorsten_verch@merck.com
- 关键词：D300 ; Direct dilution ; Low volume dispense
- 刊名：Journal of Immunological Methods
- 出版年：2017
- 出版时间：March 2017
- 年：2017
- 卷：442
- 期：Complete
- 页码：20-28
- 全文大小：1297 K
- 卷排序：442
文摘
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.