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Comparison of Compound Administration Methods in Biochemical Assays: Effects on Apparent Compound Potency Using Either Assay Ready Compound Plates or Pin tool Delivered Compounds

Smith, Thomas and Ho, Pei-I and Yue, Qing and Itkin, Zina and Macdougall, Damien and Paolucci, Michael and Hill, William and Auld, Douglas (2012) Comparison of Compound Administration Methods in Biochemical Assays: Effects on Apparent Compound Potency Using Either Assay Ready Compound Plates or Pin tool Delivered Compounds. Journal of Biomolecular Screening. ISSN 1087-0571

Abstract

Compound sample preparation and delivery is one of the most critical steps in high-throughput screening (HTS) campaigns. Historically, several methods of compound delivery to assays have been used for HTS, including pre-diluted intermediate dilution plates, “assay-ready plates” (ARP) using either pre-plated dried compound films or nanoliter DMSO spots of compounds, as well as pin tool delivered compounds. We and others have observed differences in apparent compound potency depending on the compound delivery method. To quantitatively measure compound potency differences due to the chosen delivery methods, we conducted a controlled study using a validated biochemical luciferase assay and compared potencies when compounds were delivered in either ARP (using acoustic dispensed nanoliter spots) or by pin tool. Here we compare hit rates, confirmation rates, false positive, and false negative rates between the two delivery methods using the luciferase assay. We compared polystyrene (PS) and cyclic olefin copolymer (COC) plates using both delivery methods and examined whether ARP cold stored at 4°C were superior to those stored frozen at -20°C. The data show that pin tool delivery results in more confirmed hits than pre-plated compounds, resulting in a lower false negative rate, but that this effect is minimized through the use of COC plates and by obtaining plates “just-in-time” from 4°C cold storage. We also confirmed that the choice of compound delivery method to the assay has an effect on the apparent IC50s of some compounds that confirmed in all methods. Overall, this report impacts the way HTS campaigns are utilizing assay-ready compound plates in our department.

Item Type: Article
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Additional Information: author cannot archive publisher's version/PDF
Keywords: Pin tool, high-throughput screening (HTS), acoustic dispense, compound, microtiter plate, assay ready plates, Echo
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Date Deposited: 13 Oct 2015 13:14
Last Modified: 13 Oct 2015 13:14
URI: https://oak.novartis.com/id/eprint/6959

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