WHO International Reference Reagents (IRRs) for Epidermal growth factor receptor (EGFR) variants, genomic DNA (gDNA)

With an ever-increasing number of clinically actionable genomic variants in cancer, there is an urgent need for the provision of standards to aid the definition of assay limit of detection and for the harmonisation of variant measurement in response to treatment.

In 2019, the WHO Expert Committee on Biological Standardization (ECBS) endorsed the generation of Reference Materials for clinically relevant Epidermal growth factor receptor (EGFR) variants. We planned for a series of highly characterised single-batch cancer genomic DNA (gDNA) reference materials covering multiple clinically relevant EGFR variants.

A new approach was devised in create candidate materials employing genome-editing technologies, introducing the variant of interests in the wild-type genomic context. This new approach will allow to maintain a constant germline mutational landscape and mixing of multiple materials, with minimal impact on genomic context.

This set of WHO International Reference Reagents allow:

•        Calibration of secondary standards, kits, and assays.
•        Validation of assays
•        Monitoring performance of assays.

All materials were evaluated in an international collaborative study and shown to be suitable for use in next-generation sequencing (NGS) and digital PCR (Report).

These materials may be combined to produce standards at any chosen variant percentage.

ACKNOWLEDGEMENTS

We gratefully acknowledge the significant contributions of all collaborative study participants. We express our thanks to Simon Patton of EMQN (Manchester, UK) and Klaus Pantel and Claudia Koch of ELBS for connecting us with the participants. We would also like to extend our gratitude to Paul Matejtschuk, Sara Jane Holmes, James Condron of the Markets, Manufacturing and Logistics (MML) at MHRA. We want to thank the Manufacturing Team for their invaluable work for filling and lyophilisation processes, the Inventory Team for their help on the Accelerated degradation studies and for maintaining material stocks, the Logistics Team for the support of the Collaborative study. Special thanks to Celso Neto for helping us to set up the dedicated (secure and encrypted) ShareFile Web Page and all the colleagues at MHRA that supported this work. Finally, we would like to extend our heartfelt thanks to all the team members, both present and past, whose dedication and hard work have been instrumental in the success of this project.

A full list of participants can be found in the study report.