HPLC enrichment/isolation of proteins for post-translational modification studies from complex mixtures

• Macroporous RP-HPLC for enrichment/isolation of proteins from complex mixtures.
• Non-biased protein enrichment for analyzing glycosylation.
• Sample preparation method for glycosylation analysis.

The paper describes a macroporous RP-HPLC method for separation and isolation/enrichment of proteins from complex mixtures. The method is robust and efficient; using 2.1 or 4.6 mm diameter columns provides sufficient material for subsequent proteomic analysis. The main advantage of the method is that most protein variants are isolated in the same fraction, as separation is not based on differences in isoelectric point. This is highly advantageous for studying complex mixtures and post-translational modifications. Examples related to glycosylation analysis are discussed in detail.

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