Patent ID: 7638608

Claim:
A method for separating hemagglutinin (HA) HA1 and HA2 antigens from an influenza A virus or an influenza B virus, said method comprising: applying a reduced and derivatized antigen preparation comprising solubilized HA1 and HA2 antigens and a detergent in a pH controlled solution to a Reversed-Phase High-Performance Liquid Chromatography (RP-HPLC) column, wherein a mature form of HA in an antigen preparation has been cleaved by a protease into HA1 and HA2 prior to applying the antigen preparation to the RP-HPLC column; and eluting the HA1 and HA2 antigens from the column with an ion pairing agent in an organic mobile phase, wherein the HA antigens are derivatized with an alkylating agent, wherein the elution results in separation of HA1from HA2, and wherein the elution is performed at a temperature of between about 50° C. and about 70° C.