Production of afucosylated antibodies in CHO cells by coexpression of an anti‐FUT8 intrabody

Abstract

Some effector functions prompted by immunoglobulin G (IgG) antibodies, such as antibody-dependent cell-mediated cytotoxicity (ADCC), strongly depend on the N-glycans linked to asparagine 297 of the Fc region of the protein. A single α-(1,6)fucosyltransferase (FUT8) is responsible for catalyzing the addition of an α-1,6linked fucose residue to the first GlcNAc residue of the N-linked glycans. Antibodies missing this core fucose show a significantly enhanced ADCC and increased antitumor activity, which could help reduce therapeutic dose requirement, potentially translating into reduced safety concerns and manufacturing costs.

A CHO stable pool production platform for rapid clinical development of trimeric SARS‐CoV‐2 spike subunit vaccine antigens

“…The SmT1 construct contains C-terminal FLAG-His affinity tags while all other constructs are tagless (indicated by "v3" annotation). The pTT81 ® plasmid used to generate constitutive-expression pools is based on pTT109 ® (Joubert et al, 2022) but contains only one expression cassette with a CR5 promoter for expression of a single polypeptide. For inducibleexpression pools, the pTT241 ® expression plasmid was generated from pTT81 by adding an expression cassette for CymR.…”

Strategies for controlling afucosylation in monoclonal antibodies during upstream manufacturing

“…[ 31,32 ] FUT8 can also be targeted by introducing competing glycosyltransferases, such as the overexpression of N‐Acetylglucoseaminyltransferase III, [ 33 ] or inhibitory proteins, such as the co‐expression of a membrane‐associated anti‐FUT8 intrabody within the Golgi apparatus. [ 34 ]…”

Potential targeted therapy based on deep insight into the relationship between the pulmonary microbiota and immune regulation in lung fibrosis

“…The picture depicts the core fucosylation regulates ADCC and lymphocyte information transduction about lung fibrosis. intervention of the ADCC approach (87). Furthermore, defucosylated modification may alleviate the abnormal apoptosis by natural killer cells (NK cells), which indirectly relieves the aberrant immunological response of fibrotic pathology (88).…”

“…The de-fucosylated IgG-Fc domain enhanced the induction of ADCC about 50~100-fold ( 85 , 86 ). Therefore, future studies could take CF as the artificial intervention of the ADCC approach ( 87 ). Furthermore, de-fucosylated modification may alleviate the abnormal apoptosis by natural killer cells (NK cells), which indirectly relieves the aberrant immunological response of fibrotic pathology ( 88 ).…”