Method for preparing membrane vesicles
AssigneesAnosys, Inc., Institute Curie, Institute National de la Sante et de la Recherche Medicale
InventorsOlivier Dhellin, Sebastian Amigorena, Philippe Rameau, Joël Crouzet
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Abstract
This invention relates to a method of preparing membrane vesicles from a biological sample, characterised in that it comprises at least one anion exchange and/or gel permeation chromatography treatment of the sample. The invention is used to prepare vesicles of varied origins and types, particularly from antigen presenting cells or tumoral cells. The invention also relates to the vesicles obtained in this way and their uses.
Claims15
A process of preparing membrane vesicles from the culture supernatant of a biological sample, wherein said biological sample comprises membrane vesicles produced by antigen presenting cells that have been sensitized to one or more selected antigens, said method comprising at least
- a filtration of the culture supernatant, followed by a tangential ultrafiltration to produce a biological sample enriched with membrane vesicles;
- an anion exchange chromatography treatment performed under pressure of the enriched sample followed or preceded by gel permeation chromatography of said enriched sample;
- and
- a sterilizing filtration step.
A process preparing membrane vesicles from a biological sample, wherein said process comprises at least:
- a) the culture of a population of membrane vesicles producing antigen presenting cells under conditions enabling the release of vesicles, wherein said antigen presenting cells have been sensitized to one or more selected antigens,
- b) a filtration of the culture supernatant of the cells, followed by a tangential ultrafiltration to prepare a sample enriched with membrane vesicles,
- c) an anion exchange chromatography treatment performed under pressure and a gel permeation chromatography treatment of the sample, and
- d) a sterilizing filtration step of the sample.
A process of preparing membrane vesicles, characterized in that it comprises the following steps:
- a) obtaining a population of immature dendritic cells sensitized to one or more selected antigens,
- b) culturing the dendritic cells under conditions enabling the production of membrane vesicles,
- c) treating the culture supernatant of said cells to produce a biological sample enriched with membrane vesicles by a filtration of the culture supernatant followed by a tangential ultrafiltration,
- d) purifying the membrane vesicles using a process comprising at least an anion exchange chromatography treatment performed under pressure and a gel permeation chromatography of the sample, and,
- e) a sterilising filtration step of the sample.
Process of preparing membrane vesicles from a biological sample, characterized in that it comprises:
- a) the culture of a population of membrane vesicle producing tumoral cells under conditions enabling the release of vesicles,
- b) a membrane vesicle enrichment step comprising a filtration followed by a tangential ultrafiltration,
- c) an anion exchange chromatography treatment performed under pressure and a gel permeation chromatography treatment of the sample, and
- d) a sterilizing filtration step of the sample.
Dependencies are inferred from the claim text.