Liposomes A Practical Approach

by ;
Edition: 2nd
Format: Paperback
Pub. Date: 2003-08-07
Publisher(s): Oxford University Press
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Summary

Since the first edition of "Liposomes - A Practical Approach" was published in 1990, Liposome technology has become a highly successful and rapidly developing area of preclinical and clinical research. During the 90's, a number of liposome-based drugs reached the market in the U.S. and Europe and several new aspects of liposome research have been discovered. As a consequence there has been an explosion of new procedures and techniques in the field. Reflecting both the significant changes in the field and the new discoveries, this new edition of Liposomes contains a combination of chapters from the 1st edition which have been significantly updated, and several new chapters. Each chapter is an outstanding contribution from leading experts in the field, and the book as a whole encompasses all key areas of liposome technology. The first half of the book addresses a number of essential experimental protocols such as the preparation, characterization, and storage of liposomes. Also included are methods for drug encapsulation into liposomes, modification of the liposomal surface to control drug behaviour in biological environments, long-circulating liposomes and the use of cationic liposomes as transfection vectors. The second half of the book discusses specific areas of liposome research and application, such as pH-sensitive liposomes, liposomes for diagnostic imaging, liposomal DNA vaccines, isothermic titration calorimetry of liposomes and vesicular phospholipids gels. This new edition of Liposomes is an essential tool for all experienced liposomologists and the perfect introduction for researchers new to the field.

Author Biography


Vladimir P. Torchilin is Professor and Chairman at the Department of Pharmaceutical Sciences, School of Pharmacy, Northeastern University, He gained a Ph.D.and D.Sc. in Polymer Chemistry and Chemistry of Physiologically Active Compounds, and is now a professor of Pharmaceutical Sciences. He has published over 300 papers, reviews and book chapters, 8 books and special journal issues (as author and editor). He is currently a member of the American Chemical Society, Controlled Release Society, AAPS, International Liposome Society and acts as a review editor for the Journal of Controlled Release, Associate Editor for Journal of Drug targeting, and is on the editorial boards of many other journal including Bioconjugate Chemistry and Advanced Drug Delivery Reviews.He was awarded the Lenin Prize of the USSR in Science and Technology, 1982; Member, Russian Academy of Biotechnology, 1992, and is a fellow of the American Institute for Medical and Biological Engineering, 2002.
Volkmar Weissig is an Assistant Professor of Pharmaceutics, Northeastern University, Bouve College of Health Sciences, Department of Pharmaceutical Sciences. He gained a Ph.D. in Chemistry and Sc.D. in Biochemistry and Pharmaceutical Biotechnology. Asst. He has published over 40 papers, reviews and book chapters and over 70 abstracts. He is a member of the Controlled Release Society, the International Liposome Society, the Mitochondria Research Society and the Rho Chi Honour Society.

Table of Contents

List of protocols xiii
Abbreviations xix
I General methods
1 Preparation of liposomes
3(28)
J. Lasch, V. Weissig, and M. Brandl
1 Handling and storage of lipids
3(1)
2 Preparation methods
4(19)
Hand-shaken vesicles
4(3)
Sonicated vesicles
7(2)
Freeze-dried rehydration vesicles
9(1)
Reverse-phase evaporation
10(2)
Detergent depletion
12(4)
High pressure homogenization
16(7)
3 Purification of liposomes
23(4)
Column filtration
24(1)
Purification of liposomes by centrifugation
25(2)
References
27(4)
2 Characterization of liposomes
31(48)
Nicolaas Jan Zuidam, Remco de Vrueh, and Daan J.A. Crommelin
1 Introduction
31(2)
2 Chemical analysis of liposomal components
33(17)
Spectrophotometric quantification of phospholipids
33(2)
Thin-layer chromatography of lipids
35(4)
HPLC analysis of (phospho)lipids
39(2)
HPLC analysis of cholesterol and a-tocopherol
41(1)
GLC analysis of fatty acids
42(3)
Chemical analysis of degradation of liposomal phospholipids
45(5)
3 Physical characterization of liposomes
50(22)
Determination of percentage capture
50(4)
Determination of percentage release
54(10)
Determination of entrapped volume
64(1)
Lamellarity
64(3)
Size determination of liposomes
67(5)
Zeta potential measurements based on PCS
72(1)
4 Miscellaneous methods
72(5)
Bligh and Dyer-extraction
72(2)
Sep-Pak minicolumn extraction
74(1)
Preparation of carboxyfluorescein and calcein solutions
75(2)
References
77(2)
3 Physical methods of study: differential scanning calorimetry
79(26)
Kevin M.G. Taylor and Duncan Q.M. Craig
1 Introduction
79(1)
2 Overview of techniques
79(5)
Differential scanning calorimetry
79(3)
High sensitivity differential scanning calorimetry
82(1)
Modulated temperature differential scanning calorimetry
82(2)
3 Practical aspects of making DSC measurements
84(8)
Choice of pans
84(1)
Sample size
85(1)
Calibration
85(1)
Scanning rate
86(1)
Conditions required for liposomal studies
87(2)
Practicalities of HSDSC measurements
89(1)
Practicalities of MTDSC measurements
90(2)
4 Application of DSC and related techniques to the study of liposomes
92(9)
Phase transition behaviour of liposomes
92(3)
Phase transition of multi-component systems
95(1)
Phospholipid-drug interactions
96(3)
DSC studies of liposomes as model membranes
99(1)
MTDSC of freeze-dried liposomes
99(2)
Acknowledgements
101(1)
References
101(4)
4 Fluorescence methods in liposome research
105(44)
Nejat Düzgünes, Luis A. Bagatolli, Paul Meers, Yu-Kyuong Oh, and Robert M. Straubinger
1 Fluorescence assays for liposome fusion
105(13)
The terbium/dipicolinic acid assay for intermixing of aqueous contents
105(7)
The aminonaphthalene trisulfonic acid/p-xylylene bis(pyridinium) bromide (ANTS/DPX) assay for aqueous contents mixing
112(1)
The NBD/rhodamine resonance energy transfer assay for lipid mixing
113(4)
A fluorescence assay for intermixing of inner monolayers
117(1)
2 Fluorescence assays for liposome permeability
118(1)
Release of carboxyfluorescein or calcein
118(1)
Release of the Th /DPA complex
118(1)
The release of ANTS/DPX
119(1)
3 Fluorescent detection of protein binding to phospholipid membranes
119(7)
Detection via membrane-associated fluorophores
120(3)
Detection with intrinsic protein-associated fluorophores
123(3)
4 Liposomes as pH sensors in the study of cellular interactions
126(9)
The hyranine (HPTS) assay
126(6)
The dual-fluorophore assay
132(3)
5 Two-photon excitation fluorescence microscopy to detect lipid phases in giant liposomes
135(8)
Direct visualization of lipid domain coexistence in bilayers
135(1)
Giant unilamellar liposomes
136(2)
Two-photon excitation microscopy
138(5)
Visualization of gel/fluid phase coexistence in binary phospholipid mixtures
143(1)
Acknowledgements
143(1)
References
143(6)
5 Stability, storage, and sterilization of liposomes
149(18)
Nicolaas Jan Zuidam, Ewoud van Winden, Remco de Vrueh, and Daan J.A. Crommelin
1 Introduction
149(1)
2 Prevention of chemical degradation
149(5)
Phospholipid oxidation
149(3)
Hydrolysis of phospholipids
152(2)
3 Prevention of physical changes
154(1)
4 Freeze-drying of liposomes
155(9)
Introduction
155(1)
Important factors for freeze-drying of liposomes
156(5)
Stability in the freeze-dried state
161(1)
Example of a freeze-drying protocol
162(2)
5 Preparation of sterile liposome formulations
164(1)
References
164(3)
6 Encapsulation of weakly-basic drugs, antisense oligonucleotides, and plasmid DNA within large unilamellar vesicles for drug delivery applications
167(26)
David B. Fenske, Norbert Maurer, and Pieter R. Cullis
1 Introduction
167(2)
2 Measurement of phospholipid concentration
169(2)
3 The formation of large unilamellar vesicles by extrusion through polycarbonate filters with defined pore size
171(2)
4 Accumulation of weakly-basic drugs within LUV's in response to transmembrane pH gradients
173(8)
5 Liposomal systems for the encapsulation of genetic drugs: long-circulating vectors for the systemic delivery of genes and antisense oligonucleotides
181(7)
References
188(5)
7 Surface modification of liposomes
193(38)
Vladimir P. Torchilin, Volkmar Weissig, Francis J. Martin, Timothy D. Heath, and Roger R.C. New
1 Introduction
193(1)
2 Protein and peptide attachment to the liposomal surface
194(22)
Binding of proteins and peptides to liposomes via amino groups
194(10)
Binding of proteins to liposomes via sulfhydryl groups
204(12)
3 Binding of carbohydrates and other small molecules to the liposomal surface
216(2)
4 Attachment of diagnostically significant reporter metal atoms to the liposome surface
218(9)
Conjugation of low molecular weight chelate DTPA with lipid for incorporation into the liposomal membrane
219(4)
Loading liposome-associated chelates with metals
223(4)
References
227(4)
8 Long-circulating sterically protected liposomes
231(36)
Alexander L. Klibanov, Vladimir P. Torchilin, and Samuel Zalipsky
1 Introduction
231(1)
2 Attachment of sterically protecting polymer (PEG) to liposomes
232(6)
Incorporation of PEG into liposomes
232(3)
Attachment of PEG to previously prepared liposomes
235(1)
Dependence of biodistribution on the size of long-circulating liposomes
236(1)
Tumour accumulation of long-circulating liposomes
237(1)
3 Coupling of various ligands to the distal end of liposome-grafted polymer
238(23)
Ligand attachment to hydrazide-activated PEG
243(3)
Ligand attachment to PDP-activated PEG
246(3)
Ligand attachment via COON-activated PEG
249(1)
Coupling of primary amino group-containing ligands to pNP-PEGPE-containing liposomes
250(3)
Ligand attachment by preparation of ligand-PEG-ligand conjugates and their incorporation into liposomes
253(8)
4 Detachable PEG on the liposome surface
261(1)
5 Biological factors responsible for the clearance of PEGylated liposomes
262(1)
References
263(4)
9 Liposomes in biological systems
267(22)
Jan A.A.M. Kamps and Gerrit L. Scherphof
1 Introduction
267(1)
2 Interaction of liposomes with cells
267(12)
Possible liposome-cell interaction: adsorption, (receptor-mediated) endocytosis, lipid exchange, fusion
267(2)
Markers
269(1)
Cell lines and primary cell cultures
270(3)
Isolation and culturing of liver cells
273(6)
3 Biodistribution of liposomes
279(7)
Animal models; species differences
279(1)
In vivo liposome-cell interactions
280(1)
Markers
281(1)
Liposome kinetics and tissue distribution
282(4)
Acknowledgements
286(1)
References
286(3)
10 Cationic liposomes in gene delivery
289(16)
Sean Sullivan, Yan Gong, and Jeffrey Hughes
1 Introduction
289(7)
2 Plasmid DNA isolation
296(2)
3 Quantification of plasmid DNA concentration
298(2)
4 Calculation of NSP ratio
300(1)
References
301(4)
II Selected topics and applications
11 pH-sensitive liposomes
305(14)
Regine Peschka-Süss and Rolf Schubert
1 Introduction
305(2)
2 Mechanisms of pH sensitivity
307(4)
3 Methods
311(5)
Preparation of pH-sensitive liposomes
311(3)
Characterization of pH-sensitive liposomes
314(1)
Cellular association of pH-sensitive liposomes
314(2)
References
316(3)
12 Radiolabelled liposomes for imaging and biodistribution studies
319(18)
Beth A. Goins and William T. Phillips
1 Introduction
319(1)
2 Instrumentation for tracking radiolabelled liposomes
320(3)
Gamma camera
320(2)
Positron emission tomographic camera
322(1)
Gamma well counter
322(1)
3 Methods for labelling liposomes with gamma-emitting radionuclides
323(6)
General overview
323(1)
Radionuclides available for liposome labelling methods
324(1)
Methods for labelling liposomes using radionuclides
325(4)
4 Characterization of radiolabelled liposomes
329(1)
In vitro radiolabel stability evaluation
329(1)
In vivo radiolabel evaluation
329(1)
5 Image acquisition and analysis methods
330(4)
6 Tissue biodistribution method using gamma well counter
334(1)
References
335(2)
13 Isothermic titration calorimetry (ITC)
337(16)
J. Lasch
1 Introduction
337(1)
2 Instrument and the principle of technique
337(3)
3 Computer-assisted measurement with the MCS ITC
340(2)
Making a calorimetric titration
340(2)
4 Estimation of critical micellar concentration
342(5)
Thermodynamic background
342(2)
Measurement
344(3)
5 Surfactant-bilayer membrane equilibria
347(4)
Surfactant partitioning into lipid membranes
347(2)
Solubilization of lipsomes
349(2)
References
351(2)
14 Vesicular phospholipid gels
353(20)
M. Brandl and U. Massing
1 Background
353(2)
Biopharmaceutical and technological challenges with liposomes for i.v. application
353(1)
Biopharmaceutical and technological challenges with liposomes for loco-regional administration
354(1)
Phospholipid dispersions: swelling and phases
354(1)
Mechanical stress: formulation of vesicle dispersions and vesicular phospholipid gels
354(1)
The vesicular phospholipid gel concept
355(1)
2 Preparation of vesicular phospholipid gels
355(3)
VPC's made of a single phospholipid
355(2)
VPG's made of two or more lipids
357(1)
3 Loading of VPC's with drugs
358(3)
Direct entrapment
359(1)
Passive loading
359(1)
Incorporation/association
360(1)
4 Sterilization of VPG's
361(1)
5 Characteristics of VPG's
361(4)
Morphology
361(1)
Rheology, viscosity
362(1)
Sustained release behaviour in vitro
362(3)
6 Preparation of SUV dispersions from VPG's
365(1)
7 Characterization of SUV dispersions prepared from VPG's
365(2)
Vesicle size distribution
365(1)
Encapsulation efficiency
366(1)
8 In vivo applications
367(4)
Preparation and in vitro characterization of Gemcitabine-containing VPG
368(1)
Intra-tumoral application of Gemcitabine-containing VPG
369(1)
i.v. application of Gemcitabine-containing VPG
369(2)
References
371(2)
15 Liposome-based DNA vaccines: procedures for entrapment
373(8)
Gregory Gregoriadis, Brenda McCormack, Yvonne Perrie, Andrew Bacon, Wilson Caparros-Wanderley, and Brahim Zadi
1 Introduction
373(1)
2 Materials
374(1)
3 Entrapment of plasmid DNA into liposomes by the dehydration-rehydration procedure
374(1)
Solutions
374(1)
Procedure steps
375(5)
References
380(1)
Appendix 381(8)
Index 389

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