Delivering an Automated and Integrated Approach to Combination Screening Using Acoustic-Droplet Technology

Abstract

Drug combination testing in the pharmaceutical industry has typically been driven by late-stage opportunistic strategies rather than by early testing to identify drug combinations for clinical investigation that may deliver improved efficacy. A rationale for combinations exists across a number of diseases in which pathway redundancy or resistance to therapeutics are evident. However, early assays are complicated by the absence of both assay formats representative of disease biology and robust infrastructure to screen drug combinations in a medium-throughput capacity. When applying drug combination testing studies, it may be difficult to translate a study design into the required well contents for assay plates because of the number of compounds and concentrations involved. Dispensing these plates increases in difficulty as the number of compounds and concentration points increase and compounds are subsequently rolled onto additional labware. We describe the development of a software tool, in conjunction with the use of acoustic droplet technology, as part of a compound management platform, which allows the design of an assay incorporating combinations of compounds. These enhancements to infrastructure facilitate the design and ordering of assay-ready compound combination plates and the processing of combinations data from high-content organotypic assays.

Implementation and Challenges of Direct Acoustic Dosing into Cell-Based Assays

“…The on-the-fly generation of .csv files to control the Echo has been the preferred route for all routine screening work, but Batch Managers 1 and 3 both allow the use of manually written .csv files for assay development and bespoke studies, including simple combination dosing experiments. AstraZeneca has recently developed new software tools to enable more complex combination screening, and one of these tools, Apothecary, is described in this journal edition by Cross et al 33…”

“…AstraZeneca has recently developed new software tools to enable more complex combination screening, and one of these tools, Apothecary, is described in this journal edition by Cross et al 33 Since implementing automated Echo-dosed cell-based assays in AP, assay workflows have always been split between the Integrated Echo Systems (providing compound dosing, incubation, and cell fixation) and two AutoAssay Systems 34 providing all downstream assay-processing steps (i.e., addition and wash of assay components such as permeabilizing, blocking, antibody, and detection reagent solutions). As a number of different plate-based readers have been historically required for the wide range of cell-based assays we perform, readers have also been kept on a separate automated system.…”

Why a Special Issue on Acoustic Liquid Handling?

“…This makes the technique extremely useful for dose-response experiments, as Dawes et al 22 show. The ability to send fluid from multiple wells to the same destination makes combinatorial screening much easier to set up, as reported by Chan et al, 23 Cross et al, 24 Salzer et al, 25 and others. 26,27 The opportunity to transfer low-nanoliter to mid-microliter volumes of genomic solutions, cells, and enzymes provides researchers with a means to dramatically reduce reagent and sample usage while improving results.…”

Moving Liquids with Sound: The Physics of Acoustic Droplet Ejection for Robust Laboratory Automation in Life Sciences

“…This JALA special issue does. It includes papers covering ADE application to various aspects of drug discovery (assay assembly, 20 protein crystallography, 27,28 HTS, 29,30 cell-based assays, 31 therapeutic peptides, 32 pooled compound screens, 33 and combination therapies, 34,35 as well as for (viscous) industrial enzymes, 36 genotyping, 37 clinical applications, 38,39 and even synthetic biology 40 —topics well beyond the active imaginations of Loomis and Wood nearly a century ago.…”