Synthesis, characterization, and self-assembly with plasmid DNA of a quaternary ammonium derivative of pectic galactan and its fluorescent labeling for bioimaging applications

Ramesh Chintakunta; Nitsa Buaron; Nicole Kahn; Amana Moriah; Rinat Lifshiz; Riki Goldbart; Tamar Traitel; Betty Tyler; Henry Brem; Joseph Kost

doi:10.1016/j.carbpol.2016.05.015

Synthesis, characterization, and self-assembly with plasmid DNA of a quaternary ammonium derivative of pectic galactan and its fluorescent labeling for bioimaging applications

Carbohydr Polym. 2016 Oct 5:150:308-18. doi: 10.1016/j.carbpol.2016.05.015. Epub 2016 May 10.

Authors

Affiliations

¹ Department of Chemical Engineering, Ben-Gurion University of the Negev, Israel.
² Department of Biomedical Engineering, Ben-Gurion University of the Negev, Israel.
³ Department of Neurosurgery, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
⁴ 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.
⁵ Department of Chemical Engineering, Ben-Gurion University of the Negev, Israel. Electronic address: kost@bgu.ac.il.

PMID: 27312642
DOI: 10.1016/j.carbpol.2016.05.015

Abstract

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.

Keywords: 5-DTAF; C6 rat glioma cells; CHPTAC; Cell specific carrier; Pectic galactan; Quaternized pectic galactan.

MeSH terms

Animals
Cell Line, Tumor
Chemistry Techniques, Synthetic
DNA / chemistry*
DNA / genetics*
Drug Carriers / chemical synthesis
Drug Carriers / chemistry
Fluoresceins / chemistry
Fluorescent Dyes / chemistry*
Galactans / chemical synthesis*
Galactans / chemistry
Molecular Imaging
Pectins / chemistry*
Plasmids / genetics*
Propanols / chemistry*
Quaternary Ammonium Compounds / chemistry*
Rats
Sodium Hydroxide / chemistry
Transfection
Water / chemistry

Substances

Drug Carriers
Fluoresceins
Fluorescent Dyes
Galactans
Propanols
Quaternary Ammonium Compounds
Water
5-((4,6-dichloro-1,3,5-triazin-2-yl)amino)fluorescein
Sodium Hydroxide
Pectins
DNA
(3-chloro-2-hydroxypropyl)trimethylammonium chloride