RRC ID 57003
Author Aoyama M, Tada M, Tatematsu KI, Hashii N, Sezutsu H, Ishii-Watabe A.
Title Effects of amino acid substitutions on the biological activity of anti-CD20 monoclonal antibody produced by transgenic silkworms (Bombyx mori).
Journal Biochem Biophys Res Commun
Abstract Recombinant monoclonal antibodies (mAbs) have been used in various therapeutic applications including cancer therapy. Fc-mediated effector functions play a pivotal role in the tumor-killing activities of some tumor-targeting mAbs, and Fc-engineering technologies with glyco-engineering or amino acid substitutions at the antibody Fc region have been used to enhance cytotoxic activities including antibody-dependent cellular cytotoxicity (ADCC). We previously reported that the mAbs produced using transgenic silkworms showed stronger ADCC activity and lower complement-dependent cytotoxicity (CDC) activity than mAbs derived from Chinese hamster ovary (CHO) cells due to their unique N-glycan structure (lack of core-fucose and non-reducing terminal galactose). In this study, we generated anti-CD20 mAbs with amino acid substitutions using transgenic silkworms and analyzed their biological activities to assess the effect of the combination of glyco-engineering and amino acid substitutions on the Fc-mediated function of mAbs. Three types of amino acid substitutions at the Fc region (G236A/S239D/I332E, L234A/L235A, and K326W/E333S) modified the Fc-mediated biological activities of silkworm-derived mAbs as in the case of CHO-derived mAbs, resulting in the generation of Fc-engineered mAbs with characteristic Fc-mediated functions. The combination of amino acid substitutions at the Fc region and glyco-engineering using transgenic silkworm made it possible to generate Fc-engineered mAbs with suitable Fc-mediated biological functions depending on the pharmacological mechanism of their actions. Transgenic silkworms were shown to be a promising system for the production of Fc-engineered mAbs.
Volume 503(4)
Pages 2633-2638
Published 2018-9-18
DOI 10.1016/j.bbrc.2018.08.015
PII S0006-291X(18)31688-7
PMID 30119885
MeSH Amino Acid Substitution Animals Animals, Genetically Modified Antibodies, Monoclonal, Humanized / biosynthesis Antibodies, Monoclonal, Humanized / chemistry* Antibodies, Monoclonal, Humanized / genetics Antibody-Dependent Cell Cytotoxicity* Antigens, CD20 / genetics Antigens, CD20 / immunology* Bombyx / genetics* Carbohydrate Sequence Cell Line, Tumor Fucose / chemistry Fucose / immunology Galactose / chemistry Galactose / immunology Gene Expression Humans Immunoglobulin Fc Fragments / biosynthesis Immunoglobulin Fc Fragments / chemistry* Immunoglobulin Fc Fragments / genetics Jurkat Cells Lymphocytes / cytology Lymphocytes / immunology* Polysaccharides / chemistry Polysaccharides / immunology Protein Engineering
IF 2.705
Times Cited 2