title: Purification of monoclonal antibodies to Le(y) and Le(d) carbohydrate antigens by ion-exchange and thiophilic-adsorption chromatography.
abstract: The paper deals with convenient and fast methods for purification of monoclonal antibodies (MAbs) to carbohydrate antigens Le(y) and Le(d) from the cell culture and ascite fluid by ion-exchange chromatography on S-Sepharose and salt-promoted chromatography on a "thiophilic" adsorbent. One-step procedure on S-Sepharose of MAbs to Le(y) (IgG and IgM) provides significant purification (over 90% of contaminants were removed), while a purification factor for IgM to Le(d) is much lower. Highly purified IgM to Le(d) could be obtained by a two-step purification procedure including "thiophilic-adsorption" chromatography and gel-filtration (90-98% of contaminants from the cell culture and ascite fluid were removed). The preparations of IgG and IgM retain their initial antibody activity.
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, GSP.
authors.forename: E M
authors.initials: EM
authors.lastname: Rapoport
authors.forename: L S
authors.initials: LS
authors.lastname: Zhigis
authors.forename: E V
authors.initials: EV
authors.lastname: Vlasova
authors.forename: V E
authors.initials: VE
authors.lastname: Piskarev
authors.forename: N V
authors.initials: NV
authors.lastname: Bovin
authors.forename: V P
authors.initials: VP
authors.lastname: Zubov
mesh_terms: D000327:Adsorption; D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D001201:Ascites; D002845:Chromatography / Q000379:methods; D002850:Chromatography, Gel / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D006801:Humans; D007074:Immunoglobulin G / Q000302:isolation & purification; D007075:Immunoglobulin M / Q000302:isolation & purification; D007983:Lewis Blood Group Antigens / Q000032:analysis / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C / Q000276:immunology; D008970:Molecular Weight; D012685:Sepharose
chemical_list: D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D007983:Lewis Blood Group Antigens; D012685:Sepharose