Reverse Phase Protein Arrays for High-throughput Toxicity Screening

Marlene Lemvig Pedersen, Ines Block, Markus List, Helle Christiansen, Steffen Schmidt, Jan Mollenhauer

    Research output: Contribution to conference without publisher/journalPosterResearchpeer-review

    Abstract

    High-throughput screening is extensively applied for identification of drug targets and drug discovery and recently it found entry into toxicity testing. Reverse phase protein arrays (RPPAs) are used widespread for quantification of protein markers. We reasoned that RPPAs also can be utilized beneficially in automated high-throughput toxicity testing. An advantage of using RPPAs is that, in addition to the baseline toxicity readout, they allow testing of multiple markers of toxicity, such as inflammatory responses, which do not necessarily cumulate in cell death. We used transfection of siRNAs with known killing effects as a model system to demonstrate that RPPA-based protein quantification can serve as substitute readout of cell viability, hereby reliably reflecting toxicity. In terms of automation, cell exposure, protein harvest, serial dilution and sample reformatting were performed using a robotic screening platform. Furthermore, we automated sample tracking and data analysis by developing a bundled bioinformatics tool named “MIRACLE”.
    Automation and RPPA-based viability/toxicity readouts enable rapid testing of large sample numbers, while granting the possibility for flexible consecutive quantification of additional user-defined toxicity markers.
    Original languageEnglish
    Publication date12. Oct 2015
    Publication statusPublished - 12. Oct 2015
    Event5th Global Reverse Phase Protein Array Workshop - George Mason University, Manassas, Virginia, United States
    Duration: 12. Oct 201513. Oct 2015

    Conference

    Conference5th Global Reverse Phase Protein Array Workshop
    LocationGeorge Mason University
    CountryUnited States
    CityManassas, Virginia
    Period12/10/201513/10/2015

    Fingerprint

    Protein Array Analysis
    Proteins
    Robotics
    Computational Biology
    Cell Survival
    Cell Death
    Pharmaceutical Preparations

    Cite this

    Pedersen, M. L., Block, I., List, M., Christiansen, H., Schmidt, S., & Mollenhauer, J. (2015). Reverse Phase Protein Arrays for High-throughput Toxicity Screening. Poster session presented at 5th Global Reverse Phase Protein Array Workshop, Manassas, Virginia, United States.
    Pedersen, Marlene Lemvig ; Block, Ines ; List, Markus ; Christiansen, Helle ; Schmidt, Steffen ; Mollenhauer, Jan. / Reverse Phase Protein Arrays for High-throughput Toxicity Screening. Poster session presented at 5th Global Reverse Phase Protein Array Workshop, Manassas, Virginia, United States.
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    abstract = "High-throughput screening is extensively applied for identification of drug targets and drug discovery and recently it found entry into toxicity testing. Reverse phase protein arrays (RPPAs) are used widespread for quantification of protein markers. We reasoned that RPPAs also can be utilized beneficially in automated high-throughput toxicity testing. An advantage of using RPPAs is that, in addition to the baseline toxicity readout, they allow testing of multiple markers of toxicity, such as inflammatory responses, which do not necessarily cumulate in cell death. We used transfection of siRNAs with known killing effects as a model system to demonstrate that RPPA-based protein quantification can serve as substitute readout of cell viability, hereby reliably reflecting toxicity. In terms of automation, cell exposure, protein harvest, serial dilution and sample reformatting were performed using a robotic screening platform. Furthermore, we automated sample tracking and data analysis by developing a bundled bioinformatics tool named “MIRACLE”. Automation and RPPA-based viability/toxicity readouts enable rapid testing of large sample numbers, while granting the possibility for flexible consecutive quantification of additional user-defined toxicity markers.",
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    Pedersen, ML, Block, I, List, M, Christiansen, H, Schmidt, S & Mollenhauer, J 2015, 'Reverse Phase Protein Arrays for High-throughput Toxicity Screening' 5th Global Reverse Phase Protein Array Workshop, Manassas, Virginia, United States, 12/10/2015 - 13/10/2015, .

    Reverse Phase Protein Arrays for High-throughput Toxicity Screening. / Pedersen, Marlene Lemvig; Block, Ines; List, Markus; Christiansen, Helle; Schmidt, Steffen; Mollenhauer, Jan.

    2015. Poster session presented at 5th Global Reverse Phase Protein Array Workshop, Manassas, Virginia, United States.

    Research output: Contribution to conference without publisher/journalPosterResearchpeer-review

    TY - CONF

    T1 - Reverse Phase Protein Arrays for High-throughput Toxicity Screening

    AU - Pedersen, Marlene Lemvig

    AU - Block, Ines

    AU - List, Markus

    AU - Christiansen, Helle

    AU - Schmidt, Steffen

    AU - Mollenhauer, Jan

    PY - 2015/10/12

    Y1 - 2015/10/12

    N2 - High-throughput screening is extensively applied for identification of drug targets and drug discovery and recently it found entry into toxicity testing. Reverse phase protein arrays (RPPAs) are used widespread for quantification of protein markers. We reasoned that RPPAs also can be utilized beneficially in automated high-throughput toxicity testing. An advantage of using RPPAs is that, in addition to the baseline toxicity readout, they allow testing of multiple markers of toxicity, such as inflammatory responses, which do not necessarily cumulate in cell death. We used transfection of siRNAs with known killing effects as a model system to demonstrate that RPPA-based protein quantification can serve as substitute readout of cell viability, hereby reliably reflecting toxicity. In terms of automation, cell exposure, protein harvest, serial dilution and sample reformatting were performed using a robotic screening platform. Furthermore, we automated sample tracking and data analysis by developing a bundled bioinformatics tool named “MIRACLE”. Automation and RPPA-based viability/toxicity readouts enable rapid testing of large sample numbers, while granting the possibility for flexible consecutive quantification of additional user-defined toxicity markers.

    AB - High-throughput screening is extensively applied for identification of drug targets and drug discovery and recently it found entry into toxicity testing. Reverse phase protein arrays (RPPAs) are used widespread for quantification of protein markers. We reasoned that RPPAs also can be utilized beneficially in automated high-throughput toxicity testing. An advantage of using RPPAs is that, in addition to the baseline toxicity readout, they allow testing of multiple markers of toxicity, such as inflammatory responses, which do not necessarily cumulate in cell death. We used transfection of siRNAs with known killing effects as a model system to demonstrate that RPPA-based protein quantification can serve as substitute readout of cell viability, hereby reliably reflecting toxicity. In terms of automation, cell exposure, protein harvest, serial dilution and sample reformatting were performed using a robotic screening platform. Furthermore, we automated sample tracking and data analysis by developing a bundled bioinformatics tool named “MIRACLE”. Automation and RPPA-based viability/toxicity readouts enable rapid testing of large sample numbers, while granting the possibility for flexible consecutive quantification of additional user-defined toxicity markers.

    M3 - Poster

    ER -

    Pedersen ML, Block I, List M, Christiansen H, Schmidt S, Mollenhauer J. Reverse Phase Protein Arrays for High-throughput Toxicity Screening. 2015. Poster session presented at 5th Global Reverse Phase Protein Array Workshop, Manassas, Virginia, United States.