Synthesis, characterization, and self-assembly with plasmid DNA of a quaternary ammonium derivative of pectic galactan and its fluorescent labeling for bioimaging applications.
Quaternized derivatives of pectic galactan (QPG) were synthesized by a reaction of pectic galactan (PG) with 3-chloro-2-hydroxypropyl trimethyl ammonium chloride (CHPTAC) in the presence of aqueous sodium hydroxide solution under mild reaction conditions. The results showed that the concentration of CHPTAC and NaOH has great impact on the quaternization reaction. QPG was found to interact electrostatically with plasmid DNA in aqueous solution to form complexes in globular condensed morphology in a nanometer scale size ranging from 60 to 160nm. Complexes formed with QPG fluorescently labeled with 5-DTAF (QPG-5-DTAF) were introduced to the C6 rat glioma cell line, and were found to be able to enter the cell and approach the nucleus within 24h. The results suggest that this type of modified natural polysaccharide may have an advantage as a biocompatible and biodegradable gene delivery carrier and furthermore may serve as a cell specific carrier.
Department of Chemical Engineering, Ben-Gurion University of the Negev, Israel.
Ramesh
R
Chintakunta
Department of Chemical Engineering, Ben-Gurion University of the Negev, Israel.
Nitsa
N
Buaron
Department of Chemical Engineering, Ben-Gurion University of the Negev, Israel.
Nicole
N
Kahn
Department of Biomedical Engineering, Ben-Gurion University of the Negev, Israel.
Amana
A
Moriah
Department of Chemical Engineering, Ben-Gurion University of the Negev, Israel.
Rinat
R
Lifshiz
Department of Chemical Engineering, Ben-Gurion University of the Negev, Israel.
Riki
R
Goldbart
Department of Chemical Engineering, Ben-Gurion University of the Negev, Israel.
Tamar
T
Traitel
Department of Neurosurgery, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
Betty
B
Tyler
Department of Neurosurgery, Johns Hopkins University School of Medicine, Baltimore, MD, USA; Departments of Oncology, Ophthalmology, and Biomedical Engineering, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
Henry
H
Brem
Department of Chemical Engineering, Ben-Gurion University of the Negev, Israel. Electronic address: kost@bgu.ac.il.
Joseph
J
Kost
D000818:Animals; D045744:Cell Line, Tumor; D060326:Chemistry Techniques, Synthetic; D004247:DNA / Q000737:chemistry* / Q000235:genetics*; D004337:Drug Carriers / Q000138:chemical synthesis / Q000737:chemistry; D005452:Fluoresceins / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry*; D005685:Galactans / Q000138:chemical synthesis* / Q000737:chemistry; D057054:Molecular Imaging; D010368:Pectins / Q000737:chemistry*; D010957:Plasmids / Q000235:genetics*; D020005:Propanols / Q000737:chemistry*; D000644:Quaternary Ammonium Compounds / Q000737:chemistry*; D051381:Rats; D012972:Sodium Hydroxide / Q000737:chemistry; D014162:Transfection; D014867:Water / Q000737:chemistry
D004337:Drug Carriers; D005452:Fluoresceins; D005456:Fluorescent Dyes; D005685:Galactans; D020005:Propanols; D000644:Quaternary Ammonium Compounds; D014867:Water; C039083:5-((4,6-dichloro-1,3,5-triazin-2-yl)amino)fluorescein; D012972:Sodium Hydroxide; D010368:Pectins; D004247:DNA; C496948:(3-chloro-2-hydroxypropyl)trimethylammonium chloride
5-DTAF; C6 rat glioma cells; CHPTAC; Cell specific carrier; Pectic galactan; Quaternized pectic galactan