Choice of Host Cell Line Is Essential for the Functional Glycosylation of the Fc Region of Human IgG1 Inhibitors of Influenza B Viruses

Abstract

This information is current as Viruses Human IgG1 Inhibitors of Influenza B Functional Glycosylation of the Fc Region of Choice of Host Cell Line Is Essential for the and Richard J. Pleass

Structure-based design of a highly stable, covalently-linked SARS-CoV-2 spike trimer with improved structural properties and immunogenicity

“…For practical utility as a vaccine or research reagent, a spike immunogen must be able to be efficiently produced, properly folded, and natively glycosylated in cell lines appropriate for industrial production (Blundell et al 2020; Raska et al 2010). ExpiCHO-S is a high-yield mammalian culture system derived from the commonly used industrial cell line, CHO-S, approved for Good-Manufacturing-Practice (GMP) production (He et al 2018).…”

Glycoengineering Chinese hamster ovary cells: a short history

“…The insertion of a third N-glycosylation site at the N-terminus generated highly branched and hypersialylated glycans, which are mostly absent at the Asn 297 site and when combined with further mutation of Cys residues, was able to generate a range of monomers and multimers that have tailored interactions with sialic acid-dependent receptors [ 120 ]. Some of these hypersialylated mutants were also found to bind influenza viruses with high-affinity and inhibit influenza-mediated hemagglutination [ 121 , 122 ]. While there are several fusion protein approved biotherapeutics, which mostly combine the Fc portion of IgG1 with other proteins, this section aims to highlight how backbone modifications can remove, add or multiply the functional role of glycans through the engineering of glycosylation sites and multimerisation scaffolds [ 5 ].…”

The therapeutic potential of sialylated Fc domains of human IgG

“…10,47 Approaches to enhance the sialylation of IgG have to date focussed on modifications to the known preexisting N-linked glycosylation sites. 19,40,48,49 My colleagues and I took an alternative approach to enhancing the sialic acid content of the Fc of IgG1 by adding the 18 amino acid tailpiece from IgM that contains an N-linked glycosylation site at Asn-563 to the C-terminus of the IgG1 Fc, [50][51][52][53] and into which a cysteine-toalanine substitution at Cys-575 may be introduced to prevent covalent multimerization (Figure 2). A further N-linked glycosylation site can also be introduced, if desired, to the N terminus at position Asn-221.…”

“…50 By inserting or removing in different combinations the Asn-221, Asn-297 and Asn-563 glycosylation sites, a panel of variably glycosylated Fc monomers can be generated. 51 As a result, tetravalent, octavalent, and dodecavalent Fc monomers can be made with respect to the attached terminal sialic acid (Figure 2a). Both non-covalent or covalently bonded higher ordered multimers, e.g., with possible sialic acid valences up to 72 for the hexamers depicted in Figure 2b, can then be generated from the basic Fc unit by the addition or removal of cysteines (Cys-309/Cys-575), either alone or in combination with the tailpiece Asn-563 glycan, that radically increase the available oligosaccharide combining sites (Figure 2b).…”