title: Novel Technology for Enrichment of Biomolecules from Cell-Free Body Fluids and Subsequent DNA Sizing.
abstract: Circulating cell-free DNA (ccfDNA) is a promising diagnostic tool and its size fractionation is of interest. However, kits for isolation of ccfDNA available on the market are designed for small volumes hence processing large sample volumes is laborious. We have tested a new method that enables enrichment of ccfDNA from large volumes of plasma and subsequently allows size-fractionation of isolated ccfDNA into two fractions with individually established cut-off levels of ccfDNA length. This method allows isolation of low-abundant DNA as well as separation of long and short DNA molecules. This procedure may be important e.g., in prenatal diagnostics and cancer research that have been already confirmed by our primary experiments. Here, we report the results of selective separation of 200- and 500-bp long synthetic DNA fragments spiked in plasma samples. Furthermore, we size-fractionated ccfDNA from the plasma of pregnant women and verified the prevalence of fetal ccfDNA in all fractions.
authors.affiliation: AJ Innuscreen GmbH, Berlin, Germany.
authors.forename: Vipulkumar
authors.initials: V
authors.lastname: Patel
authors.affiliation: Institute of Molecular Biomedicine, Comenius University in Bratislava, Bratislava, Slovakia.
authors.forename: Peter
authors.initials: P
authors.lastname: Celec
authors.affiliation: AJ Innuscreen GmbH, Berlin, Germany.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Grunt
authors.affiliation: Department of Tumor Biology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
authors.forename: Heidi
authors.initials: H
authors.lastname: Schwarzenbach
authors.affiliation: Analytik Jena, Jena, Germany.
authors.forename: Ingo
authors.initials: I
authors.lastname: Jenneckens
authors.affiliation: AJ Innuscreen GmbH, Berlin, Germany. timo.hillebrand@aj-innuscreen.de.
authors.forename: Timo
authors.initials: T
authors.lastname: Hillebrand
mesh_terms: D002474:Cell-Free System; D004247:DNA / Q000097:blood* / Q000235:genetics / Q000302:isolation & purification*; D005260:Female; D006801:Humans; D008967:Molecular Biology / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods; D011247:Pregnancy; D011296:Prenatal Diagnosis / Q000379:methods*; D015203:Reproducibility of Results
chemical_list: D004247:DNA
keywords: Cancer research; Cell-free DNA; DNA size-fractionation; Prenatal diagnostics