Preparation of monoclonal antibodies using the electrophoresis separation instrument, Gradiflow.
Gradiflow, a preparative electrophoresis separation device, was utilized to develop and test generic protocols for the preparation of monoclonal antibodies (MAbs) from tissue culture supernatant and ascites fluid. The charge based protocol separated the high pI antibodies from the lower isoelectric points (pI) contaminants by either moving the antibody (ascites fluid) or contaminants (tissue culture supernatant) through a polyacrylamide separation membrane. A total of 60 separations were performed with tissue culture supernatant, and a further 30 separations with ascites fluid. The Gradiflow procedure resulted in higher yields, equivalent functionality and similar purity compared with affinity chromatography antibody preparation on protein A and G. The results suggest that the Gradiflow protocols may be an alternative method of antibody preparation for these samples.
Thomas
T M
TM
Gradipore Ltd., 22 Rodborough Rd., Frenchs Forest, NSW 2086 Australia. tthomas@gradipore.com
Quindere
J
J
Thomas
D E
DE
Gee
S C
SC
Bate
I M
IM
Rylatt
D B
DB
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000302:isolation & purification*; D001202:Ascitic Fluid / Q000276:immunology; D002846:Chromatography, Affinity; D003470:Culture Media; D004586:Electrophoresis; D006801:Humans; D051379:Mice
D000911:Antibodies, Monoclonal; D003470:Culture Media