Purification of monoclonal antibodies to Le(y) and Le(d) carbohydrate antigens by ion-exchange and thiophilic-adsorption chromatography.
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.
Rapoport
E M
EM
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, GSP.
Zhigis
L S
LS
Vlasova
E V
EV
Piskarev
V E
VE
Bovin
N V
NV
Zubov
V P
VP
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
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D007983:Lewis Blood Group Antigens; D012685:Sepharose