Activity enhancement of a lung cancer-associated human monoclonal antibody HB4C5 by N-deglycosylation.
It has been known that the lung cancer-associated human monoclonal antibody HB4C5 comprises two lambda light chains of 30 and 32 kD and that the 30 kD species is exclusively responsible for the antibody activity. This study demonstrates that the two light chains were both N-glycosylated with glycosyl residues of different sizes, one of which was sensitive to neuraminidase and the other insensitive. Our unpublished data of DNA sequence for the light chain of this monoclonal antibody indicated that the light chain contains only one possible site for N-glycosylation, which located in the CDR-1. N-Deglycosylation of this monoclonal antibody under the denaturing condition resulted in the complete conversion of the two light chains into one identical polypeptide chain of 26 kD. Activity of this monoclonal antibody was found to be significantly enhanced by N-deglycosylation. All the facts described above consistently indicate that the activity of this monoclonal antibody is interfered with by the attachment of bulky glycosyl residues at the antigen binding site on the light chain. The N-deglycosylated antibody was stable under the storage conditions employed, suggesting that the glycosyl residues attached to the light chain do not play any important biological role except for interference with the antibody activity of binding antigen.
Kato
M
M
Morinaga Institute of Biological Science, Yokohama, Japan.
Mochizuki
K
K
Hashizume
S
S
Tachibana
H
H
Shirahata
S
S
Murakami
H
H
D000230:Adenocarcinoma / Q000276:immunology*; D000581:Amidohydrolases / Q000378:metabolism*; D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000378:metabolism; D000912:Antibodies, Neoplasm / Q000276:immunology* / Q000378:metabolism; D000915:Antibody Affinity / Q000187:drug effects*; D001426:Bacterial Proteins / Q000378:metabolism / Q000494:pharmacology*; D004797:Enzyme-Linked Immunosorbent Assay; D006031:Glycosylation; D006801:Humans; D007124:Immunoenzyme Techniques; D007146:Immunoglobulin lambda-Chains / Q000276:immunology / Q000378:metabolism; D007525:Isoelectric Focusing; D008175:Lung Neoplasms / Q000276:immunology*; D008969:Molecular Sequence Data; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; D011499:Protein Processing, Post-Translational; D013268:Stimulation, Chemical
D000911:Antibodies, Monoclonal; D000912:Antibodies, Neoplasm; D001426:Bacterial Proteins; D007146:Immunoglobulin lambda-Chains; D000581:Amidohydrolases; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase