Immobilization of monoclonal antibodies for affinity chromatography using a chelating peptide.
A procedure was developed for oriented immobilization of monoclonal antibodies on a solid support. The technique involves the specific oligosaccharide-directed covalent modification of the monoclonal antibody (mAb) with the chelating peptide, Lys-Gly-(His)6, in conjunction with immobilized metal ion affinity chromatography. Chelating peptide-mAb conjugates with a molar ratio of 2.2 retained full antigen binding activity. On immobilization of the modified antibodies on a nickel affinity resin, the molar antigen binding ratio was 1.4. The high antigen binding capacity is indicative of oriented immobilization providing maximum access for the antigen. The described method can be used for the preparation of high-capacity immunosorbents for affinity chromatography and it is applicable for all immunoglobulin classes.
Loetscher
P
P
F. Hoffmann-La Roche Ltd., Pharma Research, New Technologies, Basle, Switzerland.
Mottlau
L
L
Hochuli
E
E
D000911:Antibodies, Monoclonal; D002614:Chelating Agents; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D010455:Peptides / Q000737:chemistry*
D000911:Antibodies, Monoclonal; D002614:Chelating Agents; D010455:Peptides