10090714Mutated human oncogene recombinant nucleosomes as reference materials for liquid biopsyClosedCollaborative R&DInnovate UKCirculating tumour DNA (ctDNA) sequencing of plasma samples is becoming commonplace in clinical oncology practice and holds the promise to revolutionise cancer disease diagnosis and management. This promise is impeded by poor standardisation of ctDNA sequencing workflows, particularly in the pre-analytical phase. Limited control materials are currently available and published approaches often apply synthetic oligonucleotides. However, ctDNA does not circulate as free nucleic acid but predominantly as nucleosomes (DNA-histone protein complexes). Volition is the world leader in the production of recombinant-nucleosomes. These are single nucleoprotein complexes designed with the same structure in which mutated ctDNA circulates in the blood of cancer patients. These materials have the potential to fulfil the requirements of the International Liquid Biopsy Standardization Alliance for reference materials to control, monitor and standardise liquid biopsy end-to-end process from pre-analytic cell-free DNA (cfDNA) extraction through technology/assay validation and for QC of clinical sample collection and analysis. The proposed project will involve the characterisation of recombinant-nucleosome materials in terms of quantity, purity, aggregation, integrity and stability as well as their commutability with respect to native patient nucleosomes.