Purification of murine IgG monoclonal antibodies by precipitation with caprylic acid: comparison with other methods of purification.
Purification of murine IgG monoclonal antibodies from ascitic fluid by precipitation with caprylic acid was compared to that: 1) by sequential precipitation with caprylic acid and (NH4)2SO4; 2) by affinity chromatography on either antiidiotypic monoclonal antibodies or antimouse IgG xenoantibodies; and 3) by sequential (NH4)2SO4 precipitation, ion exchange chromatography and high pressure liquid chromatography (referred to as HPLC). In terms of yield of antibodies, precipitation with caprylic acid is comparable to sequential precipitation with caprylic acid and (NH4)2SO4, but superior to affinity chromatography on antibodies and to HPLC. In terms of purity of antibodies, precipitation with caprylic acid is less efficient than the other three methods. It should also be noted that precipitation with caprylic acid is associated with a reduction in the affinity of some antibodies and is not suitable to purify murine IgA and IgG3.
Temponi
M
M
Institute of Chemistry, Faculty of Medicine, University of Brescia, Italy.
Kageshita
T
T
Perosa
F
F
Ono
R
R
Okada
H
H
Ferrone
S
S
D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000302:isolation & purification; D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D000912:Antibodies, Neoplasm / Q000302:isolation & purification; D000915:Antibody Affinity; D002210:Caprylates; D011232:Chemical Precipitation; D006680:HLA Antigens / Q000276:immunology; D006801:Humans; D007074:Immunoglobulin G / Q000302:isolation & purification*; D007130:Immunoglobulin Idiotypes / Q000276:immunology; D008545:Melanoma / Q000276:immunology; D051379:Mice
D000888:Antibodies, Anti-Idiotypic; D000911:Antibodies, Monoclonal; D000912:Antibodies, Neoplasm; D002210:Caprylates; D006680:HLA Antigens; D007074:Immunoglobulin G; D007130:Immunoglobulin Idiotypes; C031492:octanoic acid