Minicircle DNA purification using a CIM® DEAE-1 monolithic support.
Minicircle DNA is a new biotechnological product with beneficial therapeutic perspectives for gene therapy because it is constituted only by the eukaryotic transcription unit. These features improve minicircle DNA safety and increase its therapeutic effect. However, being a recently developed product, there is a need to establish efficient purification methodologies, enabling the recovery of the supercoiled minicircle DNA isoform. Thus, this work describes the minicircle DNA purification using an anion exchange monolithic support. The results show that with this column it is possible to achieve a good selectivity, which allows the isolation of the supercoiled minicircle DNA isoform from impurities. Overall, this study shows a promising approach to obtain the minicircle DNA sample with adequate quality for future therapeutic applications.
CICS-UBI - Health Sciences Research Centre, Universidade da Beira Interior, Covilhã, Portugal.
Tatiana
T
Diamantino
CICS-UBI - Health Sciences Research Centre, Universidade da Beira Interior, Covilhã, Portugal.
Patrícia
P
Pereira
CICS-UBI - Health Sciences Research Centre, Universidade da Beira Interior, Covilhã, Portugal.
João A
JA
Queiroz
CICS-UBI - Health Sciences Research Centre, Universidade da Beira Interior, Covilhã, Portugal.
Ângela
Â
Sousa
CICS-UBI - Health Sciences Research Centre, Universidade da Beira Interior, Covilhã, Portugal. fani.sousa@fcsaude.ubi.pt.
Fani
F
Sousa
D000837:Anion Exchange Resins / Q000737:chemistry; D002852:Chromatography, Ion Exchange / Q000295:instrumentation / Q000379:methods*; D004278:DNA, Superhelical / Q000737:chemistry / Q000302:isolation & purification*; D006801:Humans
D000837:Anion Exchange Resins; D004278:DNA, Superhelical
Anion exchange chromatography; Minicircle DNA; Monoliths; Parental plasmids; Recombinant DNA technology