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Published on: September 13, 2013
Protocol for identifying properties of ERBB receptor antagonists using the barcoded ERBBprofiler assay
Lukša Popović1, Moritz J Rossner2, Michael C Wehr2
1Research Group Cell Signalling, Department of Psychiatry and Psychotherapy, LMU University Hospital, LMU Munich, Nussbaumstr. 7, 80336 Munich, Germany; Systasy Bioscience GmbH, Balanstr. 6, 81669 Munich, Germany.
Abstract:
The ERBBprofiler assay measures compound effects on ERBB family receptors and key downstream signaling pathways that are implicated in cancer or other complex diseases. Here, we present a protocol for identifying properties of ERBB receptor antagonists using the barcoded ERBBprofiler assay. We describe steps for in-solution transfection, cell treatment, combined processing of samples, amplification and indexing of PCRs, sequencing, and data analysis. This approach allows for the simultaneous assessment of drug effects and cell-type-dependent effects. For complete details on the use and execution of this protocol, please refer to Popović et al.1.
Insights
This study details a barcoded ERBBprofiler assay protocol for evaluating drug effects on ERBB receptors and cancer signaling pathways. The method enables simultaneous assessment of compound efficacy and cell-type-specific responses.
Area of Science:
- Biochemistry
- Molecular Biology
- Cancer Research
Background:
- The ERBB family receptors are crucial in cancer and complex diseases.
- Understanding drug effects on these pathways is vital for therapeutic development.
Purpose of the Study:
- To present a protocol for identifying properties of ERBB receptor antagonists.
- To enable simultaneous assessment of drug effects and cell-type-dependent responses.
Main Methods:
- Utilizing a barcoded ERBBprofiler assay.
- In-solution transfection, cell treatment, and combined sample processing.
- PCR amplification, indexing, sequencing, and data analysis.
Main Results:
- The protocol allows for the simultaneous assessment of drug effects.
- Cell-type-dependent effects can be evaluated concurrently.
- Identification of ERBB receptor antagonist properties is facilitated.
Conclusions:
- The barcoded ERBBprofiler assay provides a comprehensive method for evaluating ERBB pathway modulators.
- This protocol aids in understanding drug mechanisms and optimizing cancer therapies.
