Bottomley S P, Sutton B J, Gore M G
Department of Biochemistry, University of Southampton, UK.
J Immunol Methods. 1995 Jun 9;182(2):185-92. doi: 10.1016/0022-1759(95)00049-g.
Immobilised analogues of protein A have been used for affinity chromatographic separation of human IgG. Truncation of the C-terminal region of an engineered IgG-binding domain based upon the B domain from protein A, in combination with site-directed mutagenesis, has led to the generation of a number of proteins with a decreased affinity for IgG. The elution of human IgG from these proteins when immobilised onto a solid support occurs over the pH range 3.2-5.0 with 0.5 M acetate buffer. These proteins may be effective alternatives to standard protein A columns when milder elution conditions are required.
