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US 2013/0011925 A1Published application
Separation Devices and Method For Separating Phosphorylated Peptides and Proteins
AssigneeWaters Corporation
InventorsMartin Gilar, Ying-Qing Yu, Jennifer Fournier, John E. O'Gara
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Abstract
Embodiments of the present invention are directed to articles of manufacture, devices, methods and apparatus for performing liquid chromatography featuring a chromatographic sorbent having one or more pentafluorophenyl groups, wherein said one or more pentafluorophenyl groups are a bonded phase on a sorbent selected from the group comprising silica, organic polymers or hybrid organic diene material and said pentafluorophenyl groups are in a mono-, bi-, and tridentate forms.
Claims22
A chromatographic method comprising the steps of:
- loading a sample potentially comprising an analyte onto a mixed-mode chromatographic sorbent, wherein the analyte comprises a modified peptide or a modified protein and wherein the sorbent comprises a plurality of first moieties having an attractive interaction with the analyte and a plurality of second moieties having a repulsive interaction with the analyte;
- eluting the mixed-mode chromatographic sorbent in reverse phase mode, thereby separating the analyte, if present, from the sample;
- and
- eluting the mixed-mode chromatographic sorbent in ion exchange mode, thereby regenerating the mixed-mode chromatographic sorbent.
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A chromatographic method comprising the steps of:
- loading a sample potentially comprising an analyte onto a mixed-mode chromatographic sorbent, wherein the analyte comprises at least one modified peptide or modified protein and wherein the sorbent conforms to Formula 1:
- (W1)a(W2)b(W3)cSi(Z—C6FnH5-n)d Formula 1
- where W1, W2 and W3 are selected from the groups consisting of hydrogen, hydroxyl, hydroxy-aliphatic, aliphatic, oxygen, nitrogen, and silane wherein at least one of, W1, W2, and W3 separately represent terminal silane valences bonded to oxygen, nitrogen, carbon or silane atoms of a support, Z represents a aliphatic moiety and n is an integer from 1-5 and a+b+c+d=4;
- where a is 1-3;
- b+c is less than or equal to 2, and d is 1-3;
- eluting the mixed-mode chromatographic sorbent in reverse phase mode, thereby separating the analyte, if present, from the sample;
- and
- eluting the mixed-mode chromatographic sorbent in strong cation exchange mode, thereby regenerating the mixed-mode chromatographic sorbent.
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Dependencies are inferred from the claim text.