Size exclusion chromatography column technologies for analysis of CRISPR molecules
AssigneeWaters Technologies Corporation
InventorsLavelay Kizekai, Stephen Shiner, Matthew Lauber
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Abstract
The present disclosure is directed to methods for performing size exclusion chromatographic (SEC) separations. Embodiments of the present disclosure feature methods for improved separations of biomolecule analytes, such as CRISPR-related proteins, nucleotides, and ribonucleoprotein complexes, in SEC, for example, by using hydroxy-terminated polyethylene glycol surface modified stationary phase materials and/or C2/PEG surface modified column hardware.
Claims24
A method for performing size exclusion chromatography on a sample comprising at least one analyte, the at least one analyte comprising a Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-associated (Cas) protein, a CRISPR-related construct, or CRISPR-related complex, the method comprising:
- a. contacting said sample with a column chromatography device comprising a column having an interior for accepting a stationary phase, and an immobilized stationary phase within said interior of the column, wherein the column has an interior wetted surface, wherein at least a portion of its wetted surface comprises a non-ionic, hydrophilic organosilane coating, and wherein the immobilized stationary phase comprises porous particles having a surface, wherein at least some portion of the surface is modified with a hydroxy-terminated polyethylene glycol;
- b. flowing a mobile phase through the immobilized stationary phase for a period of time, the mobile phase comprising water and a buffer;
- and
- c. eluting the at least one analyte from the immobilized stationary phase in the mobile phase.
Dependencies are inferred from the claim text.