Size exclusion chromatography utilizing low concentration amino acids in size exclusion chromatography mobile phase
AssigneeWaters Technologies Corporation
InventorsAndrew Wyatt Schmudlach, Susan Rzewuski, Matthew A. Lauber, Yeliz Tunc Sarisozen, Nicole L. Lawrence, Darryl W. Brousmiche
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Abstract
The present disclosure is directed to methods for performing size exclusion chromatography. Embodiments of the present disclosure feature methods for improving separations of proteinaceous analytes in size exclusion chromatography, for example, by using low concentrations of amino acids or derivatives thereof in the mobile phase.
Claims35
A method for performing size exclusion chromatography on a sample containing at least one analyte, 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 immobilized stationary phase comprises porous particles having a surface and a diameter with a mean size distribution of between about 1 and about 20 μm;
- an average pore size from about 40 to about 3000 Å;
- and wherein said porous particles are surface modified with a hydroxy-terminated polyethylene glycol at a surface concentration from about 0.5 to about 5.0 μmoles/m2;
- b. flowing a mobile phase through the immobilized stationary phase for a period of time, the mobile phase comprising water;
- a buffer;
- and an amino acid or derivative thereof, wherein the amino acid or derivative thereof is present in the mobile phase at a concentration from about 5 to about 40 mM;
- and
- c. eluting the at least one analyte from the immobilized stationary phase in the mobile phase.
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Dependencies are inferred from the claim text.