Preparation and biological activities of anti-HER2 monoclonal antibodies with fully core-fucosylated homogeneous bi-antennary complex-type glycans

Abstract

Recently, the absence of a core-fucose residue in the N-glycan has been implicated to be important for enhancing antibody-dependent cellular cytotoxicity (ADCC) activity of immunoglobulin G monoclonal antibodies (mAbs). Here, we first prepared anti-HER2 mAbs having two core-fucosylated N-glycan chains with the single G2F, G1aF, G1bF, or G0F structure, together with those having two N-glycan chains with a single non-core-fucosylated corresponding structure for comparison, and determined their biological activities. Dissociation constants of mAbs with core-fucosylated N-glycans bound to recombinant Fcγ-receptor type IIIa variant were 10 times higher than those with the non-core-fucosylated N-glycans, regardless of core glycan structures. mAbs with the core-fucosylated N-glycans had markedly reduced ADCC activities, while those with the non-core-fucosylated N-glycans had high activities. These results indicate that the presence of a core-fucose residue in the N-glycan suppresses the binding to the Fc-receptor and the induction of ADCC of anti-HER2 mAbs.

TSTA3 facilitates esophageal squamous cell carcinoma progression through regulating fucosylation of LAMP2 and ERBB2

“…N-glycosylation inhibitors significantly reduced the binding of EGF to EGFR, and core fucosylation EGFR had a higher affinity with EGF 46 . Although fucosylation of ERBB2 has not been reported in cancer, there was evidence supporting that the antibody-dependent cellular cytotoxicity activity of the de-fucosylated antibody of ERBB2 significantly increased compared to the wild-type antibody 47 , 48 .…”

Influence of N-glycosylation on effector functions and thermal stability of glycoengineered IgG1 monoclonal antibody with homogeneous glycoforms

“…Our observations for core defucosylation were supported by many previous studies investigating both heterogeneous and homogeneous glycoforms. 12,13,22,34-42 However, no consensus has been reached regarding the effects of terminal galactosylation, with some reports describing an upregulation [36][37][38][39][42][43][44] and no effect [45][46][47] or downregulation 48 of FcγRIIIa affinity or ADCC activity. Since our results demonstrated significant statistical interaction between core fucosylation and terminal galactosylation, and are in strong agreement with previous studies utilizing mAbs with highly homogeneous glycoforms, [37][38][39]42 we hypothesize that the interpretation of results could be compromised when evaluating mixtures of different types of glycoform.…”

“…12,13,22,34-42 However, no consensus has been reached regarding the effects of terminal galactosylation, with some reports describing an upregulation [36][37][38][39][42][43][44] and no effect [45][46][47] or downregulation 48 of FcγRIIIa affinity or ADCC activity. Since our results demonstrated significant statistical interaction between core fucosylation and terminal galactosylation, and are in strong agreement with previous studies utilizing mAbs with highly homogeneous glycoforms, [37][38][39]42 we hypothesize that the interpretation of results could be compromised when evaluating mixtures of different types of glycoform. Our evaluations of the effects of terminal sialylation in FcγRIIIa binding analysis by SPR and ADCC reporter bioassay using N-glycosylated ligand agree with previous reports that have described reduction of FcγRIIIa affinity, 13,38,40,46,49,50 but found no evidence of an unchanged affinity as previously reported.…”

Generation of FX−/− and Gmds−/−CHOZN host cell lines for the production of afucosylated therapeutic antibodies

“…For some marketed oncology mAbs such as Trastuzumab, Rituximab, and Ipilimumab, the primary mechanism of actions could be attributed to antibody-dependent cellular cytotoxicity (ADCC), a process that therapeutic marketed therapeutic antibodies (mAbs) bind to specific targeted cells and recruit effector cells, which induce the apoptosis of targeted cells. [1][2][3][4] The ADCC effect is highly dependent on the ability of therapeutic mAbs to recruit effector cells such as Natural Killer (NK) cells. 5,6 The recruitment of effector cells by mAbs is determined mainly by glycan-glycan interaction between Nglycosylations on the Fc domain of mAbs and FcγRIIIa (CD16a) on effector cells.…”

“…Therapeutic mAbs have been developed and widely used for the treatment of multiple diseases, including autoimmune diseases and cancer. For some marketed oncology mAbs such as Trastuzumab, Rituximab, and Ipilimumab, the primary mechanism of actions could be attributed to antibody‐dependent cellular cytotoxicity (ADCC), a process that therapeutic marketed therapeutic antibodies (mAbs) bind to specific targeted cells and recruit effector cells, which induce the apoptosis of targeted cells 1‐4 . The ADCC effect is highly dependent on the ability of therapeutic mAbs to recruit effector cells such as Natural Killer (NK) cells 5,6 .…”