Patent ID: 7449116

Claim:
A method of protein separation from a sample comprising at least 50% of total protein being abundant proteins, the method comprising: depleting abundant proteins from a sample using immunoaffinity liquid chromatography to provide a depleted mixture of proteins; and fractionating the depleted mixture of proteins on a reversed-phase superficially porous stationary phase at a temperature of greater than or equal to 40° C., wherein the fractionating comprises a multisegmented gradient elution comprising steps of applying a first elution gradient comprising 5 to 30 weight % of organic modifier, applying a second elution gradient comprising 30 to 55 weight % of organic modifier, applying a third elution gradient comprising 55 to 100 weight % of organic modifier, applying a final elution gradient comprising 100 weight % of organic modifier; and recovering a protein from the depleted mixture of proteins at a yield of 70 to 100 weight percent of the depleted mixture of proteins.