Antibody glycosylation: impact on antibody drug characteristics and quality control

Wang, Ziyan; Zhu, Jianwei; Lu, Huili

doi:10.1007/s00253-020-10368-7

Cited by 40 publications

(24 citation statements)

References 118 publications

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“…This effect was dependent on distinct glycosylation profiles between IgA1 and IgA2 [ 42 ]. An explanation for this difference in observations may be that Hansen et al used monoclonal IgA subclasses, which are often myeloma-derived and are characterized by aberrant glycosylation profiles, while Steffen et al used IgA antibodies isolated from healthy human serum [ 40 , 42 , 68 ].…”

Section: Physiological Immune Activation: Host Defense Against Pathogensmentioning

confidence: 99%

Physiological and Pathological Inflammation Induced by Antibodies and Pentraxins

Geyer

Mes

Newling

et al. 2021

Cells

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Macrophages play a key role in induction of inflammatory responses. These inflammatory responses are mostly considered to be instigated by activation of pattern recognition receptors (PRRs) or cytokine receptors. However, recently it has become clear that also antibodies and pentraxins, which can both activate Fc receptors (FcRs), induce very powerful inflammatory responses by macrophages that can even be an order of magnitude greater than PRRs. While the physiological function of this antibody-dependent inflammation (ADI) is to counteract infections, undesired activation or over-activation of this mechanism will lead to pathology, as observed in a variety of disorders, including viral infections such as COVID-19, chronic inflammatory disorders such as Crohn’s disease, and autoimmune diseases such as rheumatoid arthritis. In this review we discuss how physiological ADI provides host defense by inducing pathogen-specific immunity, and how erroneous activation of this mechanism leads to pathology. Moreover, we will provide an overview of the currently known signaling and metabolic pathways that underlie ADI, and how these can be targeted to counteract pathological inflammation.

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Section: Physiological Immune Activation: Host Defense Against Pathogensmentioning

confidence: 99%

Physiological and Pathological Inflammation Induced by Antibodies and Pentraxins

Geyer

Mes

Newling

et al. 2021

Cells

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“…The previous study demonstrated that the mannose-type glycan of rituximab produced by P. pastoris has a higher FcγRIIIa a nity as compared to the fucosylated complex-type glycan from CHO cell line [53]. The similar comparative analysis revealed the highest FcγRIIIa a nity with the afucosylated complex-type glycan [54], and thus this enhancement is likely due to the afucosylated glycan [55]. In case of the adalimumab, the afucosylated complex-type glycan of Humira ® could have a dominant effect on the FcγRIIIa binding, while the mannnose-type glycan of adalimumab produced by A.…”

Section: Discussionmentioning

confidence: 85%

Functional production of human antibody by the filamentous fungus Aspergillus oryzae

Huynh

Morita

Sakamoto

et al. 2020

Preprint

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Background: Monoclonal antibodies (mAbs) as biopharmaceuticals take a pivotal role in the current therapeutic applications. Generally mammalian cell lines, such as those derived from Chinese hamster ovaries (CHO), are used to produce the recombinant antibody. However, there are still concerns about the high cost and the risk of pathogenic contamination when using mammalian cells. Aspergillus oryzae, a filamentous fungus recognized as a GRAS (Generally Regarded As Safe) organism, has an ability to secrete a large amount of proteins into the culture supernatant, and thus the fungus has been used as one of the cost-effective microbial hosts for heterologous protein production. Pursuing this strategy the human anti-TNFα antibody adalimumab, one of the world’s best-selling antibodies for the treatment of immune-mediated inflammatory diseases including rheumatoid arthritis, was chosen to produce the full length of mAbs by A. oryzae. Generally, N-glycosylation of the antibody affects immune effector functions such as antibody-dependent cell-mediated cytotoxicity (ADCC) via binding to the Fc receptor (FcγR) on immune cells. The CRISPR/Cas9 system was used to first delete the Aooch1 gene encoding a key enzyme for the hyper-mannosylation process in fungi to investigate the binding ability of antibody with FcγRIIIa.Results: Adalimumab was expressed in A. oryzae by the fusion protein system with α-amylase AmyB. The full-length adalimumab consisting of two heavy and two light chains was successfully produced in the culture supernatants. Among the producing strains, the highest amount of antibody was obtained from the ten-protease deletion strain (39.7 mg/L). Two-step purifications by Protein A and size-exclusion chromatography were applied to obtain the high purity sample for further analysis. The antigen-binding and TNFα neutralizing activities of the adalimumab produced by A. oryzae were comparable with those of a commercial product Humira®. No apparent binding with the FcγRIIIa was detected with the recombinant adalimumab even by altering the N-glycan structure using the Aooch1 deletion strain, which suggests only a little additional activity of immune effector functions.Conclusion: These results demonstrated an alternative low-cost platform for human antibody production by using A. oryzae, possibly offering a reasonable expenditure for patient’s welfare.

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“…The previous study demonstrated that the mannose-type glycan of rituximab produced by P. pastoris has a higher FcγRIIIa affinity as compared to the fucosylated complex-type glycan from CHO cell line [53]. The similar comparative analysis revealed the highest FcγRIIIa affinity with the afucosylated complex-type glycan [54], and thus this enhancement is likely due to the afucosylated glycan [55]. In case of the adalimumab, the afucosylated complex-type glycan of Humira ® could have a dominant effect on the FcγRIIIa binding, while the mannnose-type glycan of adalimumab produced by A. oryzae WT and Aooch1 deletion strains are inadequate.…”

Section: Discussionmentioning

confidence: 87%

Functional production of human antibody by the filamentous fungus Aspergillus oryzae

Huynh

Morita

Sakamoto

et al. 2020

Fungal Biol Biotechnol

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Background: Monoclonal antibodies (mAbs) as biopharmaceuticals take a pivotal role in the current therapeutic applications. Generally mammalian cell lines, such as those derived from Chinese hamster ovaries (CHO), are used to produce the recombinant antibody. However, there are still concerns about the high cost and the risk of pathogenic contamination when using mammalian cells. Aspergillus oryzae, a filamentous fungus recognized as a GRAS (Generally Regarded As Safe) organism, has an ability to secrete a large amount of proteins into the culture supernatant, and thus the fungus has been used as one of the cost-effective microbial hosts for heterologous protein production. Pursuing this strategy the human anti-TNFα antibody adalimumab, one of the world's best-selling antibodies for the treatment of immune-mediated inflammatory diseases including rheumatoid arthritis, was chosen to produce the full length of mAbs by A. oryzae. Generally, N-glycosylation of the antibody affects immune effector functions such as antibody-dependent cell-mediated cytotoxicity (ADCC) via binding to the Fc receptor (FcγR) on immune cells. The CRISPR/Cas9 system was used to first delete the Aooch1 gene encoding a key enzyme for the hyper-mannosylation process in fungi to investigate the binding ability of antibody with FcγRIIIa.Results: Adalimumab was expressed in A. oryzae by the fusion protein system with α-amylase AmyB. The full-length adalimumab consisting of two heavy and two light chains was successfully produced in the culture supernatants. Among the producing strains, the highest amount of antibody was obtained from the ten-protease deletion strain (39.7 mg/L). Two-step purifications by Protein A and size-exclusion chromatography were applied to obtain the high purity sample for further analysis. The antigen-binding and TNFα neutralizing activities of the adalimumab produced by A. oryzae were comparable with those of a commercial product Humira ® . No apparent binding with the FcγRIIIa was detected with the recombinant adalimumab even by altering the N-glycan structure using the Aooch1 deletion strain, which suggests only a little additional activity of immune effector functions. Conclusion:These results demonstrated an alternative low-cost platform for human antibody production by using A. oryzae, possibly offering a reasonable expenditure for patient's welfare.

show abstract

Antibody glycosylation: impact on antibody drug characteristics and quality control

Cited by 40 publications

References 118 publications

Physiological and Pathological Inflammation Induced by Antibodies and Pentraxins

Physiological and Pathological Inflammation Induced by Antibodies and Pentraxins

Functional production of human antibody by the filamentous fungus Aspergillus oryzae

Functional production of human antibody by the filamentous fungus Aspergillus oryzae

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