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Receptor-Specific Targeting with Liposomes In Vitro Based on Sterol-PEG1300 Anchors

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Abstract

Purpose

The challenge in developing liposomes to be used in active drug targeting is to design a method that can be used for modifying liposomal membranes that is applicable for a number of different specific ligands. In this study, the post insertion technique was used with activated sterol-PEG1300 anchors and was evaluated with regard to its effectiveness in active targeting in vitro. The key advantage of these anchors is that the insertion step into the liposomal membrane takes place at room temperature and is very fast.

Materials and Methods

For in vitro experiments, neuroblastoma cell lines overexpressing GD2 antigen on their surface as a target structure were chosen. This allowed the use of anti-GD2 antibodies coupled to the liposomal surface for testing of specific binding. These modified liposomes were labelled with rhodamine-PE and their cellular association was analyzed by flow cytometry.

Results

It was shown that the activated sterol-PEG1300 anchors allow specific and significant interactions of the modified liposomes with GD2 positive cells.

Conclusion

Coupling using sterol-PEG1300 anchors is both simple and rapid. It is reproducible and applicable for all ligands bearing amino groups. This method demonstrates the advantage of a ready-to-use system for the modification of pre-formed liposomes with different ligands.

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Abbreviations

Ab:

antibody

BB:

borate buffer

BSA:

bovine serum albumine

CE:

coupling efficiency

Chol:

cholesterol

EE:

encapsulation efficiency

EPC:

egg phosphatidylcholine

HBS:

HEPES buffered saline

MAL-PEG2500-DSPE:

1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[maleimido-poly(ethyleneglycol)]

NHS:

N-hydroxysuccinimide

PBS:

phosphate buffered saline

PCS:

photon correlation spectroscopy

PEG:

poly(ethyleneglycol)

PIT:

post-insertion technique

PL:

phospholipid

Rh-PE:

rhodamine-B-PE

SPIT:

sterol-based post-insertion technique

sterol-PEG1300 :

soy sterol-poly(ethyleneglycol)-1300-ether

TL:

total lipid

TLC:

thin layer chromatography

TRE:

tresyl chloride (2,2,2-trifluoroethanesulfonylchloride)

2-IT:

2-iminothiolane

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Acknowledgements

These studies were financially supported by a grant from the Mildred Scheel Foundation. The authors would like to thank Rupert Handgretinger and Stephen D Gillies for providing the antibodies and Marie Follo for proof-reading the manuscript.

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Authors and Affiliations

  1. Department of Pharmaceutical Technology and Biopharmacy, Albert-Ludwigs University, Hermann-Herder-Str. 9, 79104, Freiburg, Germany

    Markus Gantert, Thomas Steenpaß & Rolf Schubert

  2. Department of Pharmaceutical Technology and Biopharmacy, Albert-Ludwigs University, Sonnenstr. 5, 79104, Freiburg, Germany

    Felicitas Lewrick, Joanna E. Adrian & Regine Peschka-Süss

  3. Department of Pediatrics and Adolescent Medicine, Division of Pediatric Hematology and Oncology, University of Freiburg, Mathildenstr. 1, 79106, Freiburg, Germany

    Joanna E. Adrian & Jochen Rössler

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  1. Markus Gantert
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  2. Felicitas Lewrick
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Correspondence to Felicitas Lewrick.

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M. Gantert and F. Lewrick contributed equally to this publication.

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Gantert, M., Lewrick, F., Adrian, J.E. et al. Receptor-Specific Targeting with Liposomes In Vitro Based on Sterol-PEG1300 Anchors. Pharm Res 26, 529–538 (2009). https://doi.org/10.1007/s11095-008-9768-z

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  • DOI: https://doi.org/10.1007/s11095-008-9768-z

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