Show simple item record

dc.contributor.authorFarhana, Fatema Zerin
dc.contributor.authorUmer, Muhammad
dc.contributor.authorSaeed, Ayad
dc.contributor.authorPannu, Amandeep Singh
dc.contributor.authorHusaini, Sediqa
dc.contributor.authorSonar, Prashant
dc.contributor.authorFiroz, Shakhawat H
dc.contributor.authorShiddiky, Muhammad JA
dc.date.accessioned2021-05-10T00:17:45Z
dc.date.available2021-05-10T00:17:45Z
dc.date.issued2021
dc.identifier.issn0003-2654
dc.identifier.doi10.1039/d1an00345c
dc.identifier.urihttp://hdl.handle.net/10072/404207
dc.description.abstractThe quantification of global 5-methylcytosine (5mC) content has emerged as a promising approach for the diagnosis and prognosis of cancers. However, conventional methods for the global 5mC analysis require large quantities of DNA and may not be useful for liquid biopsy applications, where the amount of DNA available is limited. Herein, we report magnetic nanoparticles-assisted methylated DNA immunoprecipitation (e-MagnetoMethyl IP) coupled with electrochemical quantification of global DNA methylation. Carboxyl (-COOH) group-functionalized iron oxide nanoparticles (C-IONPs) synthesized by a novel starch-assisted gel formation method were conjugated with anti-5mC antibodies through EDC/NHS coupling (anti-5mC/C-IONPs). Anti-5mC/C-IONPs were subsequently mixed with DNA samples, in which they acted as dispersible capture agents to selectively bind 5mC residues and capture the methylated fraction of genomic DNA. The target-bound Anti-5mC/C-IONPs were magnetically separated and directly adsorbed onto the gold electrode surface using gold-DNA affinity interaction. The amount of DNA adsorbed on the electrode surface, which corresponds to the DNA methylation level in the sample, was electrochemically estimated by differential pulse voltammetric (DPV) study of an electroactive indicator [Ru(NH3)6]3+ bound to the surface-adsorbed DNA. Using a 200 ng DNA sample, the assay could successfully detect differences as low as 5% in global DNA methylation levels with high reproducibility (relative standard deviation (% RSD) = <5% for n = 3). The method could also reproducibly analyze various levels of global DNA methylation in synthetic samples as well as in cell lines. The method avoids bisulfite treatment, does not rely on enzymes for signal generation, and can detect global DNA methylation using clinically relevant quantities of sample DNA without PCR amplification. We believe that this proof-of-concept method could potentially find applications for liquid biopsy-based global DNA methylation analysis in point-of-care settings.
dc.description.peerreviewedYes
dc.description.sponsorshipThe University of Queensland ARC
dc.languageeng
dc.publisherRoyal Society of Chemistry (RSC)
dc.relation.ispartofjournalAnalyst
dc.relation.urihttp://purl.org/au-research/grants/ARC/DP190102944
dc.relation.grantIDDP190102944
dc.relation.fundersARC
dc.subject.fieldofresearchAnalytical chemistry
dc.subject.fieldofresearchOther chemical sciences
dc.subject.fieldofresearchcode3401
dc.subject.fieldofresearchcode3499
dc.titlee-MagnetoMethyl IP: a magnetic nanoparticle-mediated immunoprecipitation and electrochemical detection method for global DNA methylation.
dc.typeJournal article
dc.type.descriptionC1 - Articles
dcterms.bibliographicCitationFarhana, FZ; Umer, M; Saeed, A; Pannu, AS; Husaini, S; Sonar, P; Firoz, SH; Shiddiky, MJA, e-MagnetoMethyl IP: a magnetic nanoparticle-mediated immunoprecipitation and electrochemical detection method for global DNA methylation., Analyst, 2021
dc.date.updated2021-05-09T11:53:18Z
dc.description.versionAccepted Manuscript (AM)
gro.description.notepublicThis publication has been entered in Griffith Research Online as an advanced online version.
gro.rights.copyright© 2021 Royal Society of Chemistry. This is the author-manuscript version of this paper. Reproduced in accordance with the copyright policy of the publisher. Please refer to the journal website for access to the definitive, published version.
gro.hasfulltextFull Text
gro.griffith.authorShiddiky, Muhammad J.
gro.griffith.authorUmer, Muhammad
gro.griffith.authorPannu, Amandeep Singh S.


Files in this item

This item appears in the following Collection(s)

  • Journal articles
    Contains articles published by Griffith authors in scholarly journals.

Show simple item record