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US 2025/0164488 A1Published application
Structure and function characterization of adeno-associated virus (aav) by hdx-ms
AssigneeRegeneron Pharmaceuticals, Inc.
InventorsHaibo Qiu, Xiang Ye, Yunli Hu, Ning Li
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Abstract
The present disclosure generally pertains to methods for characterizing the structure stability and conformational dynamics of an AAV particle using Hydrogen-Deuterium Exchange Mass Spectrometry (HDX-MS).
Claims20
A method of characterizing an AAV viral particle, comprising:
- (a) incubating a sample comprising the AAV viral particle with a deuterium labeling buffer to form a labeled sample;
- (b) equilibrating a gel filtration column with greater than 4 M to about 8 M Guanidinium chloride (GndHCl);
- (c) contacting the labeled sample with the GndHCl on the equilibrated gel filtration column to form a buffer exchanged sample;
- (d) contacting the buffer exchanged sample with a hydrolyzing agent to form a digested sample;
- (e) detecting the digested sample with a liquid chromatography-mass spectrometer to determine a mass of the labeled peptides;
- and
- (f) analyzing the mass of the labeled peptides to characterize the AAV viral particle.
4 more elements
A method of characterizing at least one AAV viral particle and viral vector interaction site in a sample, comprising:
- (a) incubating a sample comprising (i) an AAV viral particle and a viral vector and a sample comprising (ii) an AAV viral particle with a deuterium labeling buffer to form labeled samples;
- (b) equilibrating a gel filtration column with Guanidinium chloride (GndHCl);
- (c) contacting the labeled samples with the GndHCl on the equilibrated gel filtration column to form buffer exchanged samples;
- (d) contacting the buffer exchanged samples with a hydrolyzing agent to form digested samples;
- (e) detecting the digested samples with a liquid chromatography-mass spectrometer to determine a mass of the labeled peptides;
- (f) analyzing the mass of the labeled peptides;
- and
- (g) analyzing hydrogen-deuterium exchange to characterize an AAV viral particle and viral vector interaction site.
5 more elements
Dependencies are inferred from the claim text.