Elution of human IgG from affinity columns containing immobilised variants of protein A.
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.
Bottomley
S P
SP
Department of Biochemistry, University of Southampton, UK.
Sutton
B J
BJ
Gore
M G
MG
D001665:Binding Sites; D002846:Chromatography, Affinity / Q000379:methods*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007074:Immunoglobulin G / Q000302:isolation & purification*; D007158:Immunologic Techniques; D016297:Mutagenesis, Site-Directed; D011485:Protein Binding; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D013205:Staphylococcal Protein A / Q000235:genetics / Q000276:immunology
D007074:Immunoglobulin G; D011994:Recombinant Proteins; D013205:Staphylococcal Protein A