Purification of monoclonal antibodies to Le(y) and Le(d) carbohydrate antigens by ion-exchange and thiophilic-adsorption chromatography

Bioseparation. 1995 Jun;5(3):141-6.

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.

MeSH terms

  • Adsorption
  • Animals
  • Antibodies, Monoclonal / isolation & purification*
  • Ascites
  • Chromatography / methods
  • Chromatography, Gel / methods
  • Chromatography, Ion Exchange / methods
  • Electrophoresis, Polyacrylamide Gel / methods
  • Humans
  • Immunoglobulin G / isolation & purification
  • Immunoglobulin M / isolation & purification
  • Lewis Blood Group Antigens / analysis
  • Lewis Blood Group Antigens / immunology*
  • Mice
  • Mice, Inbred BALB C / immunology
  • Molecular Weight
  • Sepharose

Substances

  • Antibodies, Monoclonal
  • Immunoglobulin G
  • Immunoglobulin M
  • Lewis Blood Group Antigens
  • Sepharose