title: Engineering a monomeric Fc domain modality by N-glycosylation for the half-life extension of biotherapeutics.
abstract: Human IgG is a bivalent molecule that has two identical Fab domains connected by a dimeric Fc domain. For therapeutic purposes, however, the bivalency of IgG and Fc fusion proteins could cause undesired properties. We therefore engineered the conversion of the natural dimeric Fc domain to a highly soluble monomer by introducing two Asn-linked glycans onto the hydrophobic C(H)3-C(H)3 dimer interface. The monomeric Fc (monoFc) maintained the binding affinity for neonatal Fc receptor (FcRn) in a pH-dependent manner. We solved the crystal structure of monoFc, which explains how the carbohydrates can stabilize the protein surface and provides the rationale for molecular recognition between monoFc and FcRn. The monoFc prolonged the in vivo half-life of an antibody Fab domain, and a tandem repeat of the monoFc further prolonged the half-life. This monoFc modality can be used to improve the pharmacokinetics of monomeric therapeutic proteins with an option to modulate the degree of half-life extension.
authors.affiliation: From Global Biotherapeutics Technologies, Pfizer Inc., Cambridge, Massachusetts 02140. Electronic address: tetsuya.ishino@pfizer.com.
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Ishino
authors.affiliation: Pharmacokinetics, Dynamics and Metabolism, Pfizer Inc., Andover, Massachusetts 01810.
authors.forename: Mengmeng
authors.initials: M
authors.lastname: Wang
authors.affiliation: From Global Biotherapeutics Technologies, Pfizer Inc., Cambridge, Massachusetts 02140.
authors.forename: Lidia
authors.initials: L
authors.lastname: Mosyak
authors.affiliation: From Global Biotherapeutics Technologies, Pfizer Inc., Cambridge, Massachusetts 02140.
authors.forename: Amy
authors.initials: A
authors.lastname: Tam
authors.affiliation: From Global Biotherapeutics Technologies, Pfizer Inc., Cambridge, Massachusetts 02140.
authors.forename: Weili
authors.initials: W
authors.lastname: Duan
authors.affiliation: From Global Biotherapeutics Technologies, Pfizer Inc., Cambridge, Massachusetts 02140.
authors.forename: Kristine
authors.initials: K
authors.lastname: Svenson
authors.affiliation: Pharmacokinetics, Dynamics and Metabolism, Pfizer Inc., Andover, Massachusetts 01810.
authors.forename: Alison
authors.initials: A
authors.lastname: Joyce
authors.affiliation: Pharmacokinetics, Dynamics and Metabolism, Pfizer Inc., Andover, Massachusetts 01810.
authors.forename: Denise M
authors.initials: DM
authors.lastname: O'Hara
authors.affiliation: From Global Biotherapeutics Technologies, Pfizer Inc., Cambridge, Massachusetts 02140.
authors.forename: Laura
authors.initials: L
authors.lastname: Lin
authors.affiliation: From Global Biotherapeutics Technologies, Pfizer Inc., Cambridge, Massachusetts 02140.
authors.forename: William S
authors.initials: WS
authors.lastname: Somers
authors.affiliation: From Global Biotherapeutics Technologies, Pfizer Inc., Cambridge, Massachusetts 02140.
authors.forename: Ronald
authors.initials: R
authors.lastname: Kriz
mesh_terms: D000818:Animals; D002460:Cell Line; D006031:Glycosylation; D006207:Half-Life; D015395:Histocompatibility Antigens Class I / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D011485:Protein Binding; D015202:Protein Engineering; D011961:Receptors, Fc / Q000235:genetics / Q000378:metabolism
chemical_list: D015395:Histocompatibility Antigens Class I; D007141:Immunoglobulin Fc Fragments; D011961:Receptors, Fc; C100688:Fc receptor, neonatal
keywords: Antibody Engineering; Crystal Structure; Glycosylation; Half-life Extension; N-Glycosylation; Pharmacokinetics; Protein Engineering