title: High-performance liquid chromatography of proteins on deformed nonporous agarose beads: immuno-affinity chromatography, exemplified with human growth hormone as ligand and a combination of ethylene glycol and salt for desorption of the antibodies.
abstract: An affinity chromatography column packed with nonporous agarose beads derivatized with human growth hormone via carbonyldiimidazol was used for the purification of antibodies against human growth hormone from antiserum. Desorption with 1 M sodium chloride in 60% ethylene glycol at pH 9.8 gave 100% total recovery of the antibodies, as measured by radioimmunoassay. The adsorption/desorption process is discussed in terms of hydrophobic and electrostatic interaction (these interactions may be involved in the bond between antibody and antigen in a cooperative fashion). The binding capacity of the column was estimated at about 50 micrograms of antibodies per gram sedimented agarose beads.
authors.affiliation: Institute of Biochemistry, University of Uppsala, Sweden.
authors.forename: J P
authors.initials: JP
authors.lastname: Li
authors.forename: S
authors.initials: S
authors.lastname: Hjertén
mesh_terms: D000906:Antibodies / Q000302:isolation & purification*; D000936:Antigen-Antibody Complex; D000937:Antigen-Antibody Reactions; D001665:Binding Sites; D002846:Chromatography, Affinity / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D019855:Ethylene Glycol; D005026:Ethylene Glycols / Q000737:chemistry*; D013006:Growth Hormone / Q000737:chemistry / Q000276:immunology*; D011863:Radioimmunoassay; D012685:Sepharose; D012965:Sodium Chloride / Q000737:chemistry*
chemical_list: D000906:Antibodies; D000936:Antigen-Antibody Complex; D005026:Ethylene Glycols; D012965:Sodium Chloride; D013006:Growth Hormone; D012685:Sepharose; D019855:Ethylene Glycol