Partitioning behavior of short DNA fragments in polymer/salt aqueous two-phase systems.
DNA fragment partitioning
aqueous two‐phase systems (ATPS)
cfDNA extraction
high‐throughput screening
liquid biopsy
Journal
Biotechnology journal
ISSN: 1860-7314
Titre abrégé: Biotechnol J
Pays: Germany
ID NLM: 101265833
Informations de publication
Date de publication:
Sep 2024
Sep 2024
Historique:
revised:
02
08
2024
received:
21
06
2024
accepted:
08
08
2024
medline:
9
9
2024
pubmed:
9
9
2024
entrez:
9
9
2024
Statut:
ppublish
Résumé
The development of liquid biopsy as a minimally invasive technique for tumor profiling has created a need for efficient biomarker extraction systems from body fluids. The analysis of circulating cell-free DNA (cfDNA) is especially promising, but the low amounts and high fragmentation of cfDNA found in plasma pose challenges to its isolation. While the potential of aqueous two-phase systems (ATPS) for the extraction and purification of various biomolecules has already been successfully established, there is limited literature on the applicability of these findings to short cfDNA-like fragments. This study presents the partitioning behavior of a 160 bp DNA fragment in polyethylene glycol (PEG)/salt ATPS at pH 7.4. The effect of PEG molecular weight, tie-line length, neutral salt additives, and phase volume ratio is evaluated to maximize DNA recovery. Selected ATPS containing a synthetic plasma solution spiked with human serum albumin and immunoglobulin G are tested to determine the separation of DNA fragments from the main plasma protein fraction. By adding 1.5% (w/w) NaCl to a 17.7% (w/w) PEG 400/17.3% (w/w) phosphate ATPS, 88% DNA recovery was achieved in the salt-rich bottom phase while over 99% of the protein was removed.
Identifiants
pubmed: 39246125
doi: 10.1002/biot.202400394
doi:
Substances chimiques
Polyethylene Glycols
3WJQ0SDW1A
Cell-Free Nucleic Acids
0
Sodium Chloride
451W47IQ8X
DNA
9007-49-2
Polymers
0
Salts
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
e2400394Subventions
Organisme : BioEcho Life Sciences GmbH
Informations de copyright
© 2024 The Author(s). Biotechnology Journal published by Wiley‐VCH GmbH.
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