Pharmaceutical Research

, Volume 24, Issue 5, pp 981–990

Novel Liposomal Formulation for Targeted Gene Delivery

Authors

  • Véronique Rivest
    • Molecular Endocrinology and Oncology Research CenterCentre Hospitalier de l’Université Laval (CHUL) Research Center
    • Faculty of PharmacyLaval University
  • Alix Phivilay
    • Molecular Endocrinology and Oncology Research CenterCentre Hospitalier de l’Université Laval (CHUL) Research Center
    • Faculty of PharmacyLaval University
  • Carl Julien
    • Molecular Endocrinology and Oncology Research CenterCentre Hospitalier de l’Université Laval (CHUL) Research Center
    • Faculty of PharmacyLaval University
  • Sandra Bélanger
    • Molecular Endocrinology and Oncology Research CenterCentre Hospitalier de l’Université Laval (CHUL) Research Center
    • Faculty of PharmacyLaval University
  • Cyntia Tremblay
    • Molecular Endocrinology and Oncology Research CenterCentre Hospitalier de l’Université Laval (CHUL) Research Center
    • Faculty of PharmacyLaval University
  • Vincent Émond
    • Molecular Endocrinology and Oncology Research CenterCentre Hospitalier de l’Université Laval (CHUL) Research Center
    • Faculty of PharmacyLaval University
    • Molecular Endocrinology and Oncology Research CenterCentre Hospitalier de l’Université Laval (CHUL) Research Center
    • Faculty of PharmacyLaval University
Research Paper

DOI: 10.1007/s11095-006-9224-x

Cite this article as:
Rivest, V., Phivilay, A., Julien, C. et al. Pharm Res (2007) 24: 981. doi:10.1007/s11095-006-9224-x

Abstract

Purpose

Development of a polyethylene glycol (PEG)-stabilized immunoliposome (PSIL) formulation with high DNA content suitable for in vivo intravenous administration and targeted gene delivery.

Materials and Methods

Plasmid DNA was condensed using 40% ethanol and packaged into neutral PSILs targeted to the mouse transferrin receptor using monoclonal antibodies (MAbs; clones RI7 and 8D3) attached to their PEG maleimide moieties. PSILs size was measured by quasi-elastic light scattering. The targeting capacity of the formulation was determined by transfection of mouse neuroblastoma Neuro 2A (N2A) cells with PSIL-DNA complexes conjugated with either RI7 or 8D3 MAbs.

Results

DNA encapsulation and MAb conjugation efficiencies averaged 71 ± 14% and 69 ± 5% (mean ± SD), respectively. No alteration in mean particle size (< 100 nm) or DNA leakage were found after 48 h storage in a physiological buffer, and the in vivo terminal half-life reached 23.9 h, indicating that the PSIL-DNA formulation was stable. Addition of free RI7 MAbs prevented transfection of N2A cells with PSIL-DNA complexes conjugated with either RI7 or 8D3 MAbs, confirming that the transfection was transferrin receptor-dependent.

Conclusions

The present data suggest that our new PSIL formulation combines molecular features required for targeted gene therapy including high DNA encapsulation efficiencies and vector-specific transient transfection capacity.

Key words

DNA encapsulationgene therapyliposomesmonoclonal antibodiestransferrin receptors

Abbreviations

2I

2-iminothiolane

AAV

adeno-associated virus

ATP

adenosine 5′-triphosphate

AV

adenovirus

BSA

bovine serum albumine

CMV

cytomegalovirus

[33P]-dCTP

deoxycytidine 5′-[α-33P]triphosphate

DDAB

didodecyldimethylammonium bromide

Dpm

disintegrations per minute

DSPE

distearoylphosphatidylethanolamine

EtOH

ethanol

HEPES

4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid

HSV-1

herpes simplex virus-1

LSC

liquid scintillation counting

MAbs

monoclonal antibodies

MWCO

molecular weight cut-off

N2A

neuroblastoma neuro 2A

NC

non-conjugated

3H-NSP

N-succinimidyl-[2,3-3H] propionate

PBS

phosphate buffered saline

PCR

polymerase chain reaction

pGLuc

pCMV-GLuc

PEG

polyethylene glycol

PEG2000

2,000 da polyethylene glycol

POPC

1-palmitoyl-2-oleoyl-sn-glycerol-3-phosphocholine

PSLs

PEG-stabilized liposomes

PSILs

PEG-stabilized immunoliposomes

RLU

relative light units

RT

room temperature

QELS

quasi-elastic light scattering

SEM

standard error mean

SCID

severe combined immunodeficiency

SD

standard deviation

SV40

simian virus 40

VP-SFM

virus production serum-free medium

Copyright information

© Springer Science+Business Media, LLC 2007