Method for separating peptides and proteins
AssigneeWaters Technologies 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 silane material and said pentafluorophenyl groups are in a mono-, bi-, and tridentate forms.
Claims17
1-22. (canceled)
A method for separating and isolating at least one phosphorylated peptide, phosphorylated protein, glycopeptide, or glycoprotein in a sample containing a mixture of peptides or proteins, the method comprising:
- loading the sample onto a mixed-mode chromatographic sorbent;
- eluting from the mixed-mode chromatographic sorbent in a first mode selected from the group consisting of reverse-phase mode and ion-exchange mode using a first eluent to produce a first eluate;
- eluting from the mixed-mode chromatographic sorbent in a second mode selected from the group consisting of reverse-phase mode and ion-exchange mode, the second mode being different from the first mode, using a second eluent to produce a second eluate;
- and
- isolating at least one phosphorylated peptide or phosphorylated protein analyte in the first eluate or the second eluate.
A method for separating the components of a biological sample, comprising:
- enzymatically processing the sample to form a protein digest;
- loading the processed sample onto a mixed-mode chromatographic sorbent;
- eluting from the mixed-mode chromatographic sorbent in a first mode selected from the group consisting of reverse-phase mode and ion-exchange mode using a first eluent to produce a first eluate;
- eluting from the mixed-mode chromatographic sorbent in a second mode selected from the group consisting of reverse-phase mode and ion-exchange mode, the second mode being different from the first mode, using a second eluent to produce a second eluate;
- and
- isolating at least one component of the biological sample in the first eluate or the second eluate.
Dependencies are inferred from the claim text.