Methods for making antibody fragments and compositions resulting therefrom

A method for preparing a F(ab′)2 fragment from a glycosylated antibody, said method comprising the steps of:

• (i) providing said glycosylated antibody, said glycosylated antibody comprising a hinge region, said hinge region comprising one or more protease cleavage sites located within said hinge region and one or more non-hinge regions adjacent to said hinge region, said non-hinge region(s) having one or more oligosaceharide groups attached thereto, said oligosaccharide group(s) causing said protease cleavage site(s) within said hinge region to be resistant to a protease treatment;
• (ii) exposing said glycosylated antibody to a deglycosylation composition thereby cleaving said oligosaccharide group(s) attached to said non-hinge region(s) to form a partially or wholly deglycosylated antibody having a hinge region cleavable by said protease treatment, said deglycosylation composition comprising at least one glycosidase capable of catalyzing the hydrolysis of an N-glvcosidic or O-glycosidic linkage between a sugar unit and an amino acid;
• and,
• (iii) exposing said partially or wholly deglycosylated antibody to said protease treatment to cause proteolytic cleavage of said hinge region cleavable by said protease treatment to form said F(ab′)2 fragment.