Folate-PEG-folate-graft-polyethylenimine-based gene delivery

J. M. Benns, A. Maheshwari, D. Y. Furgeson, R. I. Mahato, S. W. Kim

Research output: Contribution to journalArticle

94 Citations (Scopus)

Abstract

Folate-polyethylene glycol-folate-grafted-polyethylenimine (FPF-g-PEI) was synthesized by linking folic acid to both ends of a mono-functional PEG and then grafting to PEI. The graft ratio was determined using Beer's law by measuring the UV absorbance at 363 nm. The pH profile, diameter and shape of the carriers were determined. Transfection efficiencies were optimized in normal smooth muscle cells (SMC) and CT-26 colon adenocarcinoma cells using plasmid DNA encoding luciferase reporter gene. Free folic acid was shown to inhibit transfection with FPF-2.3g-PEI at neutral charge ratio. Relative toxicity between PEI and the modified carrier was measured using MTT colorimetric assay. Therapeutic potential of pmIFN-γ complexed with these polymeric carriers in terms of gene expression was determined at protein and mRNA levels using ELISA and RT-PCR. FPF-g-PEI was determined to have 2.3 folate-PEG-folate (FPF) linear polymers grafted to each PEI molecule. The average molecular weight was measured to be ∼33,500 Mw and the pH profile was characteristic of endosomal disruption capacity. Atomic Force Microscopy (AFM) and Dynamic Laser Light Scattering (DLLS) indicated FPF-2.3g-PEI and PEI (at 2 w/w ratio) efficiently condensed plasmid DNA resulting in oblique spheroid polyplexes with a mean diameter of ∼150 nm. FPF-2.3g-PEI was superior to PEI in terms of cytotoxicity and transfection efficiency in cancer cells. Smooth muscle cells showed no specificity for folate tethered complexes, where PEI/pLuc complexes yielded higher efficiencies.

Original languageEnglish (US)
Pages (from-to)123-139
Number of pages17
JournalJournal of Drug Targeting
Volume9
Issue number2
DOIs
StatePublished - Jan 1 2001

Fingerprint

Polyethyleneimine
Folic Acid
Transplants
Genes
Transfection
Smooth Muscle Myocytes
Plasmids
poly(ethylene glycol)-folate
Atomic Force Microscopy
DNA
Luciferases
Reporter Genes
Polymers
Colon
Adenocarcinoma
Lasers
Molecular Weight
Enzyme-Linked Immunosorbent Assay

Keywords

  • Endocytosis
  • Folic acid
  • Gene expression
  • IFN-γ
  • Luciferase
  • Polyethylenimine

ASJC Scopus subject areas

  • Pharmaceutical Science

Cite this

Folate-PEG-folate-graft-polyethylenimine-based gene delivery. / Benns, J. M.; Maheshwari, A.; Furgeson, D. Y.; Mahato, R. I.; Kim, S. W.

In: Journal of Drug Targeting, Vol. 9, No. 2, 01.01.2001, p. 123-139.

Research output: Contribution to journalArticle

Benns, J. M. ; Maheshwari, A. ; Furgeson, D. Y. ; Mahato, R. I. ; Kim, S. W. / Folate-PEG-folate-graft-polyethylenimine-based gene delivery. In: Journal of Drug Targeting. 2001 ; Vol. 9, No. 2. pp. 123-139.
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