Preparation of small amounts of sterile siRNA-liposomes with high entrapping efficiency by dual asymmetric centrifugation (DAC)

Autoren

Markus Hirsch
Vittorio Ziroli
Mark Helm
Ulrich Massing

DOI

10.1016/j.jconrel.2008.11.029

ISSN

0168-3659

Ausgabe der Veröffentlichung

1

Zeitschrift

Journal of Controlled Release

Sprache

en

Paginierung

80 - 88

Datum der Veröffentlichung

2009

Status

Published

Herausgeber

Elsevier BV

Herausgeber URL

http://dx.doi.org/10.1016/j.jconrel.2008.11.029

Datum der Datenerfassung

2018

Titel

Preparation of small amounts of sterile siRNA-liposomes with high entrapping efficiency by dual asymmetric centrifugation (DAC)

Ausgabe der Zeitschrift

135

Data source: Crossref

Abstract

Liposomal formulation of siRNA is an attractive approach for improving its delivery in vivo, shielding the RNA from nucleases and promoting tumor targeting. Here, the production of very small batch sizes of siRNA-liposomes by using the "dual asymmetric centrifugation (DAC)" technique was investigated. This new technique combines rapid and sterile liposome preparation with very high entrapping efficiencies. DAC is here presented in conjunction with a non-destructive microscale analysis based on double fluorescence labeling, which enables monitoring of siRNA integrity during the liposomal preparation. Integrity is reflected in spatial proximity of the dyes, which results in measurable fluorescence resonance energy transfer (FRET). The combination of DAC and the sensitive FRET analysis allows the handling of batch sizes down to 20 mg of conventional liposomes (CL) and sterically stabilized liposomes (SL). These were prepared in common 2 ml reaction tubes and loaded with calcein or labeled siRNA. Liposome sizes were 79+/-16 nm for CL and 109+/-9 nm for SL loaded with siRNA. Trapping efficiencies ranged from 43 to 81%, depending on batch size, enclosed compound, and liposome composition. FRET monitoring showed that the siRNA remained intact throughout DAC and that liposomal formulations protected the siRNA from nucleases. siRNA-liposomes remained stable for at least 3 months.

Addresses

Tumor Biology Center, Department of Clinical Research, Breisacher Strasse 117, Freiburg, Germany.

Autoren

Markus Hirsch
Vittorio Ziroli
Mark Helm
Ulrich Massing

DOI

10.1016/j.jconrel.2008.11.029

eISSN

1873-4995

Externe Identifier

PubMed Identifier: 19124051

Open access

false

ISSN

0168-3659

Ausgabe der Veröffentlichung

1

Zeitschrift

Journal of controlled release : official journal of the Controlled Release Society

Schlüsselwörter

Fluoresceins
RNA, Small Interfering
Liposomes
Centrifugation
Chromatography, High Pressure Liquid
Spectrometry, Fluorescence
Fluorescence Resonance Energy Transfer
Drug Compounding
Drug Stability
Sterilization
Particle Size

Sprache

eng

Medium

Print-Electronic

Online publication date

2008

Paginierung

80 - 88

Datum der Veröffentlichung

2009

Status

Published

Datum der Datenerfassung

2009

Titel

Preparation of small amounts of sterile siRNA-liposomes with high entrapping efficiency by dual asymmetric centrifugation (DAC).

Sub types

Journal Article

Ausgabe der Zeitschrift

135

Data source: Europe PubMed Central

Abstract

Liposomal formulation of siRNA is an attractive approach for improving its delivery in vivo, shielding the RNA from nucleases and promoting tumor targeting. Here, the production of very small batch sizes of siRNA-liposomes by using the "dual asymmetric centrifugation (DAC)" technique was investigated. This new technique combines rapid and sterile liposome preparation with very high entrapping efficiencies. DAC is here presented in conjunction with a non-destructive microscale analysis based on double fluorescence labeling, which enables monitoring of siRNA integrity during the liposomal preparation. Integrity is reflected in spatial proximity of the dyes, which results in measurable fluorescence resonance energy transfer (FRET). The combination of DAC and the sensitive FRET analysis allows the handling of batch sizes down to 20 mg of conventional liposomes (CL) and sterically stabilized liposomes (SL). These were prepared in common 2 ml reaction tubes and loaded with calcein or labeled siRNA. Liposome sizes were 79+/-16 nm for CL and 109+/-9 nm for SL loaded with siRNA. Trapping efficiencies ranged from 43 to 81%, depending on batch size, enclosed compound, and liposome composition. FRET monitoring showed that the siRNA remained intact throughout DAC and that liposomal formulations protected the siRNA from nucleases. siRNA-liposomes remained stable for at least 3 months.

Date of acceptance

2008

Autoren

Markus Hirsch
Vittorio Ziroli
Mark Helm
Ulrich Massing

Autoren-URL

https://www.ncbi.nlm.nih.gov/pubmed/19124051

DOI

10.1016/j.jconrel.2008.11.029

eISSN

1873-4995

Ausgabe der Veröffentlichung

1

Zeitschrift

J Control Release

Schlüsselwörter

Centrifugation
Chromatography, High Pressure Liquid
Drug Compounding
Drug Stability
Fluoresceins
Fluorescence Resonance Energy Transfer
Liposomes
Particle Size
RNA, Small Interfering
Spectrometry, Fluorescence
Sterilization

Sprache

eng

Country

Netherlands

Paginierung

80 - 88

PII

S0168-3659(08)00775-X

Datum der Veröffentlichung

2009

Status

Published

Datum, an dem der Datensatz öffentlich gemacht wurde

2009

Titel

Preparation of small amounts of sterile siRNA-liposomes with high entrapping efficiency by dual asymmetric centrifugation (DAC).

Sub types

Journal Article

Ausgabe der Zeitschrift

135

Data source: PubMed

Preparation of small amounts of sterile siRNA-liposomes with high entrapping efficiency by dual asymmetric centrifugation (DAC)

Tools