High-throughput chromatography screening for extracellular vesicles
AssigneeLonza Sales AG
InventorsMichael Doherty, Aaron Noyes, Andrew Wood
Life of this patent
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Abstract
The present disclosure relates to high-throughput screening methods for identifying one or more chromatography operational parameters (e.g., binding parameters) and/or reagents for purifying EVs (e.g., exosomes) from a sample using chromatography. Also disclosed herein are methods for improving one or more aspects of EV (e.g., exosome) purification, e.g., improving EV yield, increasing EV ligand density, and/or reducing impurity recovery.
Claims17
A method of purifying an extracellular vesicle (EV) comprising a prostaglandin F2 receptor negative regulator (PTGFRN) protein or a fragment thereof expressed in or on the EV from a sample, the method comprising
- (i) contacting the sample to an ion-exchange chromatography resin, a size exclusion chromatography resin, an ion-exchange chromatography medium, or a size exclusion chromatography medium, wherein the resin or medium is not a mixed mode chromatography resin or medium, under a plurality of chromatography operational parameters,
- (ii) collecting a fraction comprising the EV from step (i), and
- (iii) determining an EV yield, impurity recovery, and/or EV ligand density from step (ii),
- wherein the contacting of the sample to the resin or medium occurs in a microplate or in miniature columns, and
- wherein the PTGFRN protein or fragment thereof anchors a payload or an exogenous biologically active molecule to the EV.
A method of identifying one or more chromatography operational parameters suitable for purifying an extracellular vesicle (EV) comprising a prostaglandin F2 receptor negative regulator (PTGFRN) protein or a fragment thereof expressed in or on the EV from a sample comprising the EV and an impurity with a chromatography, the method comprising:
- (i) contacting the sample to an ion-exchange chromatography resin, a size exclusion chromatography resin, an ion-exchange chromatography medium, or a size exclusion medium, wherein the resin or medium is not a mixed mode chromatography resin or medium, under a plurality of chromatography operational parameters,
- (ii) collecting a fraction comprising the EV from step (i),
- (iii) determining an EV yield, impurity recovery, and/or EV ligand density from step (ii), and
- (iv) identifying the one or more chromatography operational parameters, wherein the one or more identified chromatography operational parameters are suitable for purifying the EV from the sample if (a) the EV yield is increased, (b) the impurity recovery is reduced, and/or (c) the EV ligand density is increased, as compared to a corresponding value in the sample prior to step (i),
- wherein the contacting of the sample to the resin or medium occurs in a microplate or in miniature columns, and
- wherein the PTGFRN protein or fragment thereof anchors a payload or an exogenous biologically active molecule to the EV.
Dependencies are inferred from the claim text.