Water-soluble lipopolymer for gene delivery

Sang Oh Han, Ram I. Mahato, Sung Wan Kim

Research output: Contribution to journalArticle

201 Citations (Scopus)

Abstract

The use of biocompatible polymeric gene carriers may overcome the current problems associated with viral vectors in safety, immunogenicity, and mutagenesis. Nontoxic water-soluble lipopolymer (WSLP), poly{(ethylenimine)-co-[N-(2-aminoethyl) ethyleneimin]-co-N-(N-cholesteryloxycarbonyl-(2-aminoethyl)-ethylenimine)} was synthesized using branched poly(ethylenimine) (PEI, mw 1800) and cholesteryl chloroformate. Following synthesis and purification, the structure and molecular weight of WSLP were confirmed by 1H NMR and MADI-TOF mass spectrometry, respectively. The percentage of cholesterol conjugated to PEI was about 47%, and the average molecular weight of WSLP was approximately 2000 Da. WSLP/pDNA complexes were prepared at different N/P (nitrogen atoms of WSLP/phosphate of plasmid DNA) ratios and characterized in terms of particle size, ζ potential, osmolarity, surface morphology, and cytotoxicity. WSLP condensed plasmid DNA when N/P ratio reached 2.5/1 and no free DNA was detected at N/P ratio of 5/1 and above, as determined by agarose gel electrophoresis. The mean particle size was in the range of 25.9 to 148.5 nm and was dependent on N/P ratios. Atomic force microscopy (AFM) showed complete condensation of plasmid DNA with spherical particles of ∼50 nm in diameter. WSLP/pDNA complexes or WSLP itself were nontoxic to CT-26 colon adenocarcinoma and 293 T human embryonic kidney transformed cells when formulated at the N/P ratio of 10/1 and below as determined by MTT assay. In contrast, PEI25000/pDNA complexes were toxic to these cells. Erythrocytes aggregated when incubated with PEI25000/pCMV-Luc complexes at high DNA concentrations, but there was little aggregation with WSLP/pCMV-Luc complexes. WSLP/pCMV-Luc complexes demonstrated higher transfection efficiency in both CT-26 and 293 T cells compared to PEI25000- or PEI1800-based formulations. WSLP/pCMV-Luc complexes are nontoxic and showed enhanced in vitro transfection. Thus, WSLP will be a suitable carrier for in vivo gene delivery.

Original languageEnglish (US)
Pages (from-to)337-345
Number of pages9
JournalBioconjugate Chemistry
Volume12
Issue number3
DOIs
StatePublished - May 1 2001

Fingerprint

Genes
Water
DNA
Polyetherimides
Plasmids
Particle Size
poly(ethylenimine)-co-(N-(2-aminoethyl) ethyleneimin)-co-N-(N-cholesteryloxycarbonyl-(2-aminoethyl)ethylenimine)
Transfection
Molecular weight
Particle size
Molecular Weight
Mutagenesis
T-cells
Cholesterol
Agar Gel Electrophoresis
Atomic Force Microscopy
Poisons
Cytotoxicity
Electrophoresis
Purification

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Biomedical Engineering
  • Pharmacology
  • Pharmaceutical Science
  • Organic Chemistry

Cite this

Water-soluble lipopolymer for gene delivery. / Han, Sang Oh; Mahato, Ram I.; Kim, Sung Wan.

In: Bioconjugate Chemistry, Vol. 12, No. 3, 01.05.2001, p. 337-345.

Research output: Contribution to journalArticle

Han, Sang Oh ; Mahato, Ram I. ; Kim, Sung Wan. / Water-soluble lipopolymer for gene delivery. In: Bioconjugate Chemistry. 2001 ; Vol. 12, No. 3. pp. 337-345.
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