Thiophilic adsorption: rapid purification of F(ab)2 and Fc fragments of IgG1 antibodies from murine ascitic fluid.
A thiophilic adsorption method has been developed for rapid purification and separation of mouse F(ab)2 and Fc fragments obtained after proteolytic digestion of IgG1 monoclonal antibodies. Partially purified Mabs were digested with papain. Thiophilic chromatography was performed using stepwise elution with decreasing concentrations of ammonium sulphate. Most contaminating proteins did not react with the thiophilic adsorbent, and chromatography efficiently resolved the F(ab)2 and Fc fragments, as judged by electrophoresis. Fractions containing the F(ab)2 fragments retained about 90% of the total antibody activity loaded onto the column.
Yurov
G K
GK
All-Russia Research Institute of Agricultural Biotechnology, Moscow.
Neugodova
G L
GL
Verkhovsky
O A
OA
Naroditsky
B S
BS
D000256:Adenoviridae / Q000276:immunology; D000327:Adsorption; D000818:Animals; D000914:Antibodies, Viral / Q000032:analysis / Q000302:isolation & purification; D001201:Ascites; D004797:Enzyme-Linked Immunosorbent Assay; D007140:Immunoglobulin Fab Fragments / Q000032:analysis / Q000302:isolation & purification*; D007141:Immunoglobulin Fc Fragments / Q000032:analysis / Q000302:isolation & purification*; D051379:Mice; D008807:Mice, Inbred BALB C; D010206:Papain; D010449:Peptide Mapping
D000914:Antibodies, Viral; D007140:Immunoglobulin Fab Fragments; D007141:Immunoglobulin Fc Fragments; D010206:Papain