Tandem purification of IgM monoclonal antibodies from mouse ascites fluids by anion-exchange and gel fast protein liquid chromatography.

Abstract

A tandem chromatographic procedure was used to isolate rapidly mouse IgM monoclonal antibodies. Mouse ascites fluids containing IgM monoclonal antibodies were first chromatographed on an anion-exchange Mono Q column connected to a fast protein liquid chromatography system. The IgM-rich fractions from the Mono Q column were then injected on a gel filtration Superose 6 column equilibrated with a low-ionic strength buffer and eluted with a high-ionic strength buffer. Assessment of the purity of isolated IgM monoclonal antibodies was performed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis together with a Coomassie Brillant Blue R 250 staining technique. Assessment of the immunoreactivity of isolated IgM monoclonal antibodies was performed by a enzyme linked immunosorbent assay using a solid phase adsorbed antigen against which IgM monoclonal antibodies were directed. The chromatographic procedure described provides a new method for the rapid purification of mouse IgM monoclonal antibodies to a high degree of purity and in a immunoreactive state.

• Authors: P Clezardin, G Bougro, J L McGregor
• Journal: Journal of chromatography (J Chromatogr) Vol. 354 Pg. 425-33 (Feb 28 1986) Netherlands
• PMID: 3700533 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antibodies, Monoclonal
• Immunoglobulin M

Topics

• Animals
• Antibodies, Monoclonal (isolation & purification)
• Ascitic Fluid (immunology)
• Chromatography, Gel
• Chromatography, Ion Exchange
• Enzyme-Linked Immunosorbent Assay
• Immunoglobulin M (immunology, isolation & purification)
• Mice