Methods for isolation of cell-free DNA using an anti-double-stranded DNA antibody
Provided herein are compositions and methods for isolating cell-free nucleic acid, e.g., cell-free DNA, from a sample. In particular embodiments, provided herein are compositions and methods using anti-dsDNA antibodies for isolating cell-free DNA from a sample, and for providing a sample of isolated cell-free DNA, e.g., for a nucleic acid assay. In particular embodiments, the technology relates to providing cell-free DNA from a maternal sample that is enriched for fetal cell-free fetal DNA.
1. A method for preparing cell-free DNA from a blood plasma sample from a human subject, comprising:
a) mixing the plasma sample with a solution comprising an exogenous anti-double-stranded DNA (anti-dsDNA) antibody that has reactivity to both single-stranded and double-stranded DNA to form a solution comprising antibody-DNA complexes comprising the anti-dsDNA antibody and cell-free DNA;
b) combining the solution comprising the antibody-DNA complexes with an antibody-binding reagent to produce bound antibody-DNA complexes; and
c) separating the bound antibody-DNA complexes from the solution comprising the antibody-DNA complexes to provide isolated cell-free DNA.
2. The method of claim 1 , further comprising a step d) of releasing isolated cell-free DNA from the bound antibody-DNA complexes.
3. The method of claim 1 , further comprising assaying the isolated cell-free DNA by a method that comprises adding isolated cell-free DNA to a reaction mixture.
4. The method of claim 3 , wherein adding isolated cell-free DNA to a reaction mixture comprises adding isolated cell-free DNA that has been released from an antibody-DNA complex to the reaction mixture.
5. The method of claim 3 , wherein adding isolated cell-free DNA to a reaction mixture comprises adding the bound antibody-DNA complexes to the reaction mixture.
6. The method of claim 3 , wherein the reaction mixture comprises at least one nucleic acid-modifying enzyme selected from a nucleic acid polymerase, a nuclease, and a ligase.
7. The method of claim 1 , wherein the human subject is a pregnant subject, and wherein the isolated cell-free DNA comprises cell-free fetal DNA.
8. The method of claim 1 , wherein the isolated cell-free DNA comprises a plurality of dsDNA fragments having lengths of between 50 and 200 bp and having a size distribution comprising peaks at about 143 bp and 166 bp.
9. The method of claim 1 , wherein the human subject is suspected of having a tumor.