Preparation of immuno-stimulating complexes (ISCOMs) by ether injection

Pham, Hoang Lam; Shaw, P. Nicholas; Davies, Nigel

doi:10.1016/j.ijpharm.2005.11.011

Cited by 19 publications

(11 citation statements)

References 18 publications

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“…If required, the organic solvent can be removed readily, and the buffer exchanged by dialysis or by gel filtration chromatography. Metabolites, drugs and other components such as peptides and proteins can be incorporated or encapsulated, thus accounting for the considerable interest in this technique for the preparation of lipid nanoparticles (Pham et al, 2006;Schubert and Müller-Goymann, 2003;Zarif, 2002;Miclea et al, 2007), resulting in the creation of a number of patents by the pharmaceutical industry.…”

Section: Introductionmentioning

confidence: 99%

Cholesterol microcrystals and cochleate cylinders: Attachment of pyolysin oligomers and domain 4

Harris

Lewis

Baik

et al. 2011

Journal of Structural Biology

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Section: Introductionmentioning

confidence: 99%

Cholesterol microcrystals and cochleate cylinders: Attachment of pyolysin oligomers and domain 4

Harris

Lewis

Baik

et al. 2011

Journal of Structural Biology

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“…For systems prepared with saponin fractions 1 and 2 (unidentified saponins), no evidence of ISCOM formation was noted using TEM when these fractions were combined with phospholipid and cholesterol at a ratio of 3:5:2 by the ether injection technique [25]. Instead, bilayer structures (characteristic of liposomes) and long helices were observed as the dominant structures (Fig.…”

Section: Preparation Of Iscoms From Purified Fractionsmentioning

confidence: 98%

“…The purified fractions of Quil A, as isolated by semipreparative HPLC and freeze-dried, were used for the preparation of ISCOMs or related colloidal structures by our previously published method of ether injection using a mass ratio of 5: 3: 2 of phosphatidylcholine (PC): Quil A: cholesterol (Chol) [25]. This ratio was found to form homogenous ISCOMs when crude Quil A was used as the saponin whereas other ratios resulted in more heterogeneous preparations in which ISCOMs typically existed together with other colloidal structures.…”

Section: Preparation Of Iscom Matrices From Purified Saponinsmentioning

confidence: 99%

“…Finally, the resulting preparation was allowed to equilibrate at room temperature with stirring for 3 days prior to characterisation of the resulting dispersion. An equilibration time of 3 days was chosen based on our previous study which indicated no differences in the colloidal structures formed from PC: Quil A: Chol, between samples taken at Day 3, Day 7, and even after 2 months of storage for a range of PC: Quil A: Chol ratios [25].…”

Section: Preparation Of Iscom Matrices From Purified Saponinsmentioning

confidence: 99%

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Saponins from Quillaja saponaria Molina: Isolation, Characterization and Ability to Form Immuno Stimulatory Complexes (ISCOMs)

Pham¹,

Ross²,

McGeary³

et al. 2006

CDD

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ISCOMs have received much attention as vaccine adjuvants due to their immunostimulatory effects. They are colloidal particles typically comprised of phospholipids, cholesterol and Quil A, a crude mixture of saponins extracted from the bark of Quillaja saponaria Molina. We have previously shown that ISCOMs can be prepared by ether injection wherein an ether solution of phospholipids and cholesterol in a mass ratio of 5:2 is injected into a solution of Quil A at a mass ratio of 7 lipids: 3 Quil A. The aim of this study was firstly to isolate and characterise discrete fractions of Quil A and secondly to investigate which of these fractions were able to form ISCOMs by the method of ether injection. Six fractions of Quil A were isolated by semi-preparative reverse phase high performance liquid chromatography (RP-HPLC) and characterised by analytical HPLC, liquid chromatography tandem mass spectrometry (LC-MS) and the qualitative Liebermann-Burchard and Molisch tests for triterpenoids and carbohydrates respectively. ISCOMs were subsequently prepared from the isolated fractions by the method of ether injection and the resulting preparations characterized by photon correlation spectroscopy (PCS) and negative stain transmission electron microscopy (TEM). The molecular weights of the major compounds in the fractions ranged from approximately 1200 to approximately 2300 Da; all fractions tested positive for triterpenoids and saccharides and four of the fractions were identified as QS-7, QS-17, QS-18 and QS-21 by analysis (LC-MS and analytical HPLC). Injection of ether solutions of lipids into aqueous solutions of QS-17, QS-18 or QS-21 all resulted in homogeneous ISCOM dispersions. The combination of lipids and QS-7 by ether injection produced lamellae and liposomes as the prominent structures and a minor amount of ISCOMs. The remaining two hydrophilic, low molecular weight fractions of Quil A did not produce ISCOMs, instead liposomes and helical structures predominated in the samples.

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“…It has been shown that ISCOMs display high adjuvant activity against a broad range of bacterial and viral antigens [1-5]. However, the side effects of ISCOMs are their toxicity, caused by the presence of the hemolytic saponins of Q. saponaria [2,6], a consistently insufficient adjuvant activity, and the absence of a satisfactory method of preparing them for industrial applications [6-9]. The development of the ISCOMATRIX™ adjuvant, based on purified saponin fractions from Quil A substantially overcomes these shortcomings [1,10].…”

Section: Introductionmentioning

confidence: 99%

Tubular immunostimulating complex based on cucumarioside A2-2 and monogalactosyldiacylglycerol from marine macrophytes

et al. 2011

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Background: There is an urgent need to develop safe and effective adjuvants for the new generation of subunit vaccines. We developed the tubular immunostimulating complex (TI-complex) as a new nanoparticulate antigen delivery system. The morphology and composition of TI-complexes principally differ from the known vesicular immunostimulating complexes (ISCOMs). However, methodology for the preparation of TI-complexes has suffered a number of shortcomings. The aim of the present work was to obtain an antigen carrier consisting of triterpene glycosides from Cucumaria japonica, cholesterol, and monogalactosyldiacylglycerol from marine macrophytes with reproducible properties and high adjuvant activity.

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Preparation of immuno-stimulating complexes (ISCOMs) by ether injection

Cited by 19 publications

References 18 publications

Cholesterol microcrystals and cochleate cylinders: Attachment of pyolysin oligomers and domain 4

Cholesterol microcrystals and cochleate cylinders: Attachment of pyolysin oligomers and domain 4

Saponins from Quillaja saponaria Molina: Isolation, Characterization and Ability to Form Immuno Stimulatory Complexes (ISCOMs)

Tubular immunostimulating complex based on cucumarioside A2-2 and monogalactosyldiacylglycerol from marine macrophytes

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