Simplified preparation of rabbit Fab fragments.
Papain attached to solid-phase CH-Sepharose 4B was used to digest rabbit IgG. Protein A-Sepharose CL-4B was used to remove undigested IgG and Fc fragments. Pure Fab fragments free of IgG, Fc fragments and papain were readily obtained by this procedure with a yield of about 75%. Polyacrylamide gel electrophoresis of the Fab in the presence of sodium dodecyl sulphate gave a single band under both reducing and non-reducing conditions. The molecular weight of the Fab determined by sedimentation equilibrium was 49,200. Unlike the IgG, the Fab obtained did not form precipitin lines when used in immunoelectrophoresis.
D000818:Animals; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D007124:Immunoenzyme Techniques; D007140:Immunoglobulin Fab Fragments / Q000032:analysis / Q000302:isolation & purification*; D007074:Immunoglobulin G / Q000032:analysis; D051379:Mice; D008970:Molecular Weight; D009498:Neurotoxins / Q000276:immunology; D011817:Rabbits; D012910:Snake Venoms / Q000276:immunology
D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G; D009498:Neurotoxins; D012910:Snake Venoms