Pepsin digestion of a mouse monoclonal antibody of IgG1 class formed F(ab')(2) fragments in which the light chains as well as the heavy chains were truncated.
In the preparation of F(ab')(2) fragments of monoclonal antibodies (mAbs) of IgG class, heavy (H) chains are truncated by pepsin and light (L) chains are remained intact. However, F(ab')(2) fragments formed by pepsin-digestion of a mouse mAb PM373, which was of the IgG1 class and raised against human prostate specific antigen (PSA), indicated that the L chains of 31 kDa were cleaved into 23-kDa fragments as well as the cleavage of H chains of 50 kDa into 28-kDa fragments. On the other hand, F(ab')(2) fragments formed by digesting the mAb by cathepsin D showed that the L chains were intact and the H chains were truncated. The immunoreactivities against PSA of the F(ab')(2) fragments containing the intact L chains and those containing the truncated L chains were almost the same as that of the parental mAb, suggesting that the truncation of the L chains does not affect the interaction of the mAb with its specific antigen.
Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Sakyo-ku, 606-8502, Kyoto, Japan. inouye@kais.kyoto-u.ac.jp
K
K
Inouye
S
S
Ohnaka
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000378:metabolism*; D002402:Cathepsin D / Q000737:chemistry / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000737:chemistry / Q000378:metabolism*; D007074:Immunoglobulin G / Q000737:chemistry / Q000378:metabolism*; D007147:Immunoglobulin Light Chains / Q000737:chemistry / Q000378:metabolism*; D051379:Mice; D010434:Pepsin A / Q000737:chemistry / Q000378:metabolism*; D017430:Prostate-Specific Antigen / Q000276:immunology
D000911:Antibodies, Monoclonal; D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G; D007147:Immunoglobulin Light Chains; D017430:Prostate-Specific Antigen; D010434:Pepsin A; D002402:Cathepsin D