Method for the enrichment of circulating tumor dna
AssigneeBelgian Volition SPRL
InventorsJacob Vincent MICALLEF, Marielle HERZOG, Mark Edward ECCLESTON
Life of this patent
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Abstract
The invention relates to the use of histone binding agents for detecting, isolating and/or purifying cell free nucleosomes of tumor originor circulating tumor DNA from a biological sample. The invention also relates to methods and kits using said histone binding agents.
Claims27
(canceled)
A method for isolating circulating cell free nucleosomes of tumor origin from a biological sample by affinity purification wherein said method comprises the steps of:
- (i) contacting the sample with a histone H3.1 and/or H3.2 and/or H3t binding agent;
- (ii) isolating bound nucleosomes from the sample;
- and
- (iii) analysing the isolated nucleosomes and/or associated DNA.
A method for isolating purified circulating tumor DNA (ctDNA) from a biological sample, wherein said method comprises the steps of:
- (i) isolating circulating cell free nucleosomes containing histone H3.1 and/or H3.2 and/or H3t;
- (ii) extracting DNA from the nucleosome sample produced in step (i);
- and
- (iii) analysing the extracted DNA.
An immunoassay method for detecting an epigenetic epitope of tumor derived circulating nucleosomes in a biological sample, wherein said method comprises the steps of:
- (i) contacting the sample with a histone H3.1 and/or H3.2 and/or H3t binding agent;
- (ii) contacting the nucleosomes or sample with a second binding agent which binds to said epitope;
- (iii) detecting and/or quantifying the binding of said second binding agent to said epitope;
- and
- (iv) using the presence or degree of such binding as a measure of the presence of the particular epitope of tumor derived nucleosomes in the sample.
An immunoassay method for detecting an epigenetic epitope of tumor derived circulating nucleosomes in a biological sample wherein said method comprises the steps of:
- (i) contacting the sample with a first binding agent which binds to said epitope;
- (ii) contacting the nucleosomes or sample with a histone H3.1 and/or H3.2 and/or H3t binding agent;
- (iii) detecting and/or quantifying the binding of said histone H3.1 and/or H3.2 and/or H3t binding agent to nucleosomes in the sample;
- and
- (iv) using the presence or degree of such binding as a measure of the presence of the particular epitope of tumor derived nucleosomes in the sample.
A method of diagnosing cancer which comprises the step of detecting circulating cell free nucleosome associated histone variant H3.1 and/or H3.2 and/or H3t in a biological sample obtained from a human or animal subject.
(canceled)
Dependencies are inferred from the claim text.