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Publications | Labcyte Inc.

featuring the Echo® Acoustic Technology
PUBLICATIONS and other Scientific Content

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TITLES and AUTHORS Year Link PDF + Abstract
  • Precision Cancer Medicine in the Acoustic Dispensing Era: Ex Vivo Primary Cell Drug Sensitivity Testing Evgeny Kulesskiy, Jani Saarela, Laura Turunen, and Krister Wennerberg Publication: Journal of Laboratory Automation (JALA) Special Issue 2016 abstract
    Related Applications:
    ADME Tox, 
    Related Product(s):
    Echo 525 for High-volume Transfers, Echo 55X for Screening, 

    Cancer therapy is increasingly becoming individualized, but there are also big gaps between the molecular knowledge of individual cancers we can generate today and what can be applied in the clinic. In an attempt to bridge this knowledge gap between cancer genetic and molecular profiling and clinically useful information, an individualized systems medicine program has been established at the Institute for Molecular Medicine Finland (FIMM), University of Helsinki, and the Helsinki University Hospital. Central to this program is drug sensitivity and resistance testing (DSRT), in which responses of primary cancer cells to a comprehensive clinical oncology and signal transduction drug collection are monitored. The drug sensitivity information is used with molecular profiling to establish hypotheses on individual cancer-selective targeting drug combinations and their predictive biomarkers, which can be explored in the clinic. Here, we describe how acoustic droplet ejection is enabling DSRT in our cancer individualized systems medicine program to (1) generate consistent but configurable assay-ready plates and determine how this affects data quality, (2) flexibly prepare drug combination testing plates, (3) dispense reagents and cells to the assay plates, and (4) perform ultra-miniaturized follow up assays on the cells from DSRT plates.

    Publication / Type:
    Journal of Laboratory Automation (JALA) Special Issue
    Related Subject:
    automated biology, statistics, informatics and software, compound management, acoustic droplet ejection
    Link:
    http://jla.sagepub.com/content/21/1/27.full.pdf+html
  • Revisiting Cell Screening to Find New Cancer Drug Combos Mark Ratner Publication: IN VIVO: Business & Medicine Report 2016 PDF abstract
    Related Applications:
    HTS and Secondary Screening, 

    A handful of academic groups in Europe and the US are using high-throughput (HT) ex vivo screening to test the sensitivity of individual patient tumors to hundreds of combinations of cancer drugs - a strategy that in earlier iterations failed to predict response to therapy. HT screening is used routinely in drug discovery. Thanks to advances in nanoliter-scale sample handling and computational biology, evidence is slowly building the potential of HT approaches to identify novel combination therapies, first in leukemia and hopefully for treating other cancers.

    Publication / Type:
    IN VIVO: Business & Medicine Report
    Related Subject:
    IN VIVO, HT, high throughput and drug discovery
  • Individualized Medicine vs. Precision Medicine Joe Olechno Publication: DDNEWS 2016 abstract
    Related Applications:
    Personalized Medicine, 
    Related Product(s):
    Echo 55X for Screening, Access Laboratory Workstation, 

    There are people who will die of cancer this week even though there are drugs that could help them.

    At the same time, hundreds of patients will undergo cancer chemotherapy that, while debilitating and expensive, will not cure them of their disease. While cancer is a formidable foe, there is a way to improve patient care and prognosis immediately.

    Researchers in Finland, Sweden and Spain have modified ex-vivo testing of cancer cells with significant results. Their approach is far more “personalized” than traditional precision medicine. Their results are striking. They provide a missing link between genomics-based mutation determination and clinical efficacy. Precision medicine (also referred to as “personalized medicine” or PM) appears to many patients, doctors and researchers to be a golden highway from disease identification to cure. The idea of interrogating the genome of a particular cancer to determine its Achilles heel is intuitively satisfying and understandable. There are, however, significant problems. First, PM is neither personalized nor precise. PM strives to identify the appropriate biomarker (usually a DNA mutation but proteins, peptides and metabolites can stand as biomarkers as well) to categorize the patient as a member of a specific group of patients. The patient is treated with a drug that has shown positive results on previous members of the group. In other words, personalized medicine is actually population-based medicine.

    Publication / Type:
    DDNEWS
    Related Subject:
    precision medicine, cancer treatment,
    Link:
    http://www.labcyte.com/resources/articles/individualized-vs-precision-medicine-olechno
  • Identification of b-hematin inhibitors in the MMV Malaria Box Fong K., Sandlin R. and Wright D. Publication: International Journal for Parasitology: Drugs and Drug Resistance, Volume 5, Issue 3, December 2015 2015 abstract
    Related Product(s):
    Echo 55X for Screening and OMICS, 

    The Malaria Box, assembled by the Medicines for Malaria Venture, is a set of 400 structurally diverse, commercially available compounds with demonstrated activity against blood-stage Plasmodium falciparum. The compounds are a representative subset of the 20,000 in vitro antimalarials identified from the high-throughput screening efforts of St. Jude Children's Research Hospital (TN, USA), Novartis and GlaxoSmithKline. In addition, a small set of active compounds from commercially available libraries was added to this group, but it has not previously been published. Elucidation of the biochemical pathways on which these compounds act is a major challenge; therefore, access to these compounds has been made available free of charge to the investigator community. Here, the Malaria Box compounds were tested for activity against the formation of b-hematin, a synthetic form of the heme detoxification biomineral, hemozoin. Further, the mechanism of action of these compounds within the malaria parasite was explored. Ten of the Malaria Box compounds demonstrated significant inhibition of b-hematin formation. In this assay, doseeresponse data revealed IC50 values ranging from 8.7 to 22.7 mM for these hits, each of which is more potent than chloroquine (a known inhibitor of hemozoin formation). The in vitro antimalarial activity of these ten hits was confirmed in cultures of the chloroquine sensitive D6 strain of the parasite resulting in IC50 values of 135e2165 nM, followed by testing in the multidrug resistant strain, C235. Cultures of P. falciparum (D6) were then examined for their heme distribution following treatment with nine of the commercially available confirmed compounds, seven of which disrupted the hemozoin pathway.

    Publication / Type:
    International Journal for Parasitology: Drugs and Drug Resistance, Volume 5, Issue 3, December 2015
    Related Subject:
    b-hematin inhibitors, MMV malaria
    Link:
    http://www.sciencedirect.com/science/article/pii/S2211320715000111
  • Compound Transfer by Acoustic Droplet Ejection Promotes Quality and Efficiency in Ultra-High-Throughput Screening Campaigns Dawes T., Turincio R., Jones S., Rodriguez R., Gadiagellan D., Thana P., Clark K., Gustafson A., Orr Publication: Journal of Laboratory Automation (JALA) Special Issue 2015 abstract
    Related Applications:
    HTS and Secondary Screening, 
    Related Product(s):
    Echo 55X for Screening, Echo 55X for Screening and OMICS, 

    Acoustic droplet ejection (ADE) as a means of transferring library compounds has had a dramatic impact on the way in which high-throughput screening campaigns are conducted in many laboratories. Two Labcyte Echo ADE liquid handlers form the core of the compound transfer operation in our 1536-well based ultra-high-throughput screening (uHTS) system. Use of these instruments has promoted flexibility in compound formatting in addition to minimizing waste and eliminating compound carryover. We describe the use of ADE for the generation of assay-ready plates for primary screening as well as for follow-up dose-response evaluations. Custom software has enabled us to harness the information generated by the ADE instrumentation. Compound transfer via ADE also contributes to the screening process outside of the uHTS system. A second fully automated ADE-based system has been used to augment the capacity of the uHTS system as well as to permit efficient use of previously picked compound aliquots for secondary assay evaluations. Essential to the utility of ADE in the high-throughput screening process is the high quality of the resulting data. Examples of data generated at various stages of high-throughput screening campaigns are provided. Advantages and disadvantages of the use of ADE in high-throughput screening are discussed.

    Publication / Type:
    Journal of Laboratory Automation (JALA) Special Issue
    Related Subject:
    high-throughput screening, acoustic droplet ejection, assay-ready plates
    Link:
    http://jla.sagepub.com/content/21/1/64.full.pdf+html
  • Acoustic Dispensing Preserves the Potency of Therapeutic Peptides throughout the Entire Drug Discovery Workflow Naylor J, Rossi A, and Hornigold D Publication: J Lab Autom. 2015 May 22. DOI: 10.1177/2211068215587915 2015 PDF abstract

    Routine peptide structure-activity relationship screening requires the serial dilution of peptides to produce full concentration response curves. Established tip-based protocols involve multiple tip changes and high exposure to plasticware. In the case of peptides, this becomes a challenge, since peptides can adsorb to plastic, resulting in an observed loss of potency. Various methods can be employed to prevent peptide loss during compound handling, such as the inclusion of bovine serum albumin or solvents in assay buffer and the siliconization of plasticware, yet protein binding remains unpredictable. The degree of variation by which peptides will adhere to plasticware can confuse results and cause inaccuracies in potency predictions. We evaluated acoustic non-contact methods for peptide serial dilution and compared it with traditional tip-based methods, on the effect on potency curves for glucagon-like peptide-1 and glucagon peptide analogues. The current study demonstrates the benefits of non-contact dispensing for high-density microplate assay preparation of peptides using nanoliter droplets across our entire drug discovery workflow, from in vitro high-throughput screening to drug exposure determinations from in vivo samples.

    Publication / Type:
    J Lab Autom. 2015 May 22. DOI: 10.1177/2211068215587915
    Related Subject:
    acoustic droplet ejection, cAMP, EC50, GLP-1, glucagon
  • Delivering an Automated and Integrated Approach to Combination Screening Using Acoustic-Droplet Technology Cross K, Craggs R, Swift D, Stiaram A, Daya S, Roberts M, Hawley S, Owen P, and Isherwood V Publication: J Lab Autom. 2015 Apr 2. DOI: 10.1177/2211068215579163 2015 PDF abstract
    Related Applications:
    HTS and Secondary Screening, 
    Related Product(s):
    Echo 55X for Screening, Echo 55X for Screening and OMICS, 

    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.

    Publication / Type:
    J Lab Autom. 2015 Apr 2. DOI: 10.1177/2211068215579163
    Related Subject:
    laboratory informatics, combinations, acoustic droplet technology, apothecary, assay-ready plate, high-content screening, sample management
  • Miniaturization of an HTRF® Epigenetic Assay with the Labcyte® Echo® Liquid Handler and the BMG PHERAstar FS® Plate Reader Edwards B, Lesnick J, Peters C and Tang N Publication: POSTER 2015 PDF abstract

    Epigenetics continues to emerge as an important target class for drug discovery and cancer research. As programs scale to evaluate many new targets related to epigenetic expression, new tools and techniques are required to enable efficient and reproducible highthroughput epigenetic screening. Echo liquid handlers can transfer compounds, samples, and reagents in sub-microliter volumes to high density assay formats using only acoustic energy - no contact or tips required. This eliminates tip costs and reduces the risk of reagent carryover. The PHERAstar FS multi-mode plate reader, with the highest sensitivity and lowest read time of assays in high density plate formats, is a perfect complement to enable an unparalleled solution for cost-effective, high-throughput epigenetic screening. Using the HTRF EPIgeneous™ Methyltransferase kit from Cisbio, we developed a miniaturized methyltransferase assay that can be easily adapted to automation and increased throughput, while maintaining high data quality. HTRF assays are typically performed at volumes of about 20 µL in a 384-well low volume plate. However, with the nanoliter dispense increments of the Echo liquid handler, assay volumes can be reduced significantly while maintaining data quality. In this study we were able to reduce a methyltransferase assay to a 2 µL final volume with excellent results.

    Publication / Type:
    POSTER
    Related Subject:
    HTRF® Epigenetic Assay Labcyte® Echo® Liquid Handler BMG PHERAstar FS® Plate
  • qPCRTag Analysis - A High Throughput, Real Time PCR Assay for Sc2.0 Genotyping Mitchell L, Phillips N, Lafont A, Martin J, Cutting R, and Boeke J Publication: J. Vis. Exp. (99), e52941, doi:10.3791/52941 (2015) 2015 abstract
    Related Applications:
    Genotyping, 
    Related Product(s):
    Echo 55X for OMICS, Echo 55X for Screening and OMICS, 

    The Synthetic Yeast Genome Project (Sc2.0) aims to build 16 designer yeast chromosomes and combine them into a single yeast cell. To date one synthetic chromosome, synIII1, and one synthetic chromosome arm, synIXR2, have been constructed and their in vivo function validated in the absence of the corresponding wild type chromosomes. An important design feature of Sc2.0 chromosomes is the introduction of PCRTags, which are short, re-coded sequences within open reading frames (ORFs) that enable differentiation of synthetic chromosomes from their wild type counterparts. PCRTag primers anneal selectively to either synthetic or wild type chromosomes and the presence/absence of each type of DNA can be tested using a simple PCR assay. The standard readout of the PCRTag assay is to assess presence/absence of amplicons by agarose gel electrophoresis However, with an average PCRTag amplicon density of one per 1.5 kb and a genome size of ~12 Mb, the completed Sc2.0 genome will encode roughly 8,000 PCRTags. To improve throughput, we have developed a real time PCR-based detection assay for PCRTag genotyping that we call qPCRTag analysis. The workflow specifies 500 nl reactions in a 1,536 multiwell plate, allowing us to test up to 768 PCRTags with both synthetic and wild type primer pairs in a single experiment.

    Publication / Type:
    J. Vis. Exp. (99), e52941, doi:10.3791/52941 (2015)
    Related Subject:
    Echo® Liquid Handler for OMICS , Echo® Liquid Handler for Screening and OMICS
    Link:
    http://www.jove.com/video/52941/qpcrtag-analysis-high-throughput-real-time-pcr-assay-for-sc20
  • Low-Cost, High-Throughput Sequencing of DNA Assemblies Using a Highly Multiplexed Nextera Process E Shapland, et. al. Publication: ACS Synth. Biol., Article ASAP DOI: 10.1021/sb500362n 2015 abstract
    Related Applications:
    Sequencing, DNA Assembly, 
    Related Product(s):
    Echo 55X for Screening and OMICS, 

    In recent years, next-generation sequencing (NGS) technology has greatly reduced the cost of sequencing whole genomes, whereas the cost of sequence verification of plasmids via Sanger sequencing has remained high. Consequently, industrial-scale strain engineers either limit the number of designs or take short cuts in quality control. Here, we show that over 4000 plasmids can be completely sequenced in one Illumina MiSeq run for less than $3 each (15× coverage), which is a 20-fold reduction over using Sanger sequencing (2× coverage). We reduced the volume of the Nextera tagmentation reaction by 100-fold and developed an automated workflow to prepare thousands of samples for sequencing. We also developed software to track the samples and associated sequence data and to rapidly identify correctly assembled constructs having the fewest defects. As DNA synthesis and assembly become a centralized commodity, this NGS quality control (QC) process will be essential to groups operating high-throughput pipelines for DNA construction.

    Publication / Type:
    ACS Synth. Biol., Article ASAP DOI: 10.1021/sb500362n
    Related Subject:
    synthetic biology; next-generation sequencing; NGS; high throughput, DNA Assembly, Sequencing, Echo® Liquid Handler for Screening and OMICS
    Link:
    http://pubs.acs.org/doi/abs/10.1021/sb500362n
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