Cell Free Remodeling of Glycosylation of Antibodies

Abstract

The N-glycosylation profile of a monoclonal antibody (mAb) is a critical quality attribute in relation to its therapeutic application. The control of this profile during biomanufacture is difficult because of the multiple parameters that affect the glycosylation metabolism within the cell and the environment in which the cell is grown. One of the approaches that can be used to produce a preferred glycan profile or a single glycoform is through chemoenzymatic remodeling during the isolation of a mAb. Here we describe protocols that can be utilized to produce preferred glycoforms that include galactosylated, agalactosylated, or sialylated glycoforms following isolation of a mAb. Methods for analysis and assignment of structures of the samples following glycoengineering are also described. Chemoenzymatic modeling of mAb glycans has the potential for scale-up and to be introduced into biomanufacturing of mAbs with higher specific therapeutic activities.

First identification of canine adenovirus 1 in mink and bioinformatics analysis of its 100 K protein

“…Studies have shown that the newly developed equipment can perform high-speed and accurate research on abnormal glycosylation (Bangarh et al, 2023 ). Although glycosylation accounts for 2–3% of the total mass of IgG antibodies, its therapeutic application is mainly attributed to the N-glycosylation profile of a monoclonal antibody (mAb) (Mota et al, 2022 ). The Mink CAdV-1 100 K protein has only one glycosylation site.…”

Identification and characterization of a β-1,4 Galactosidase fromElizabethkingia meningosepticaand its application on living cell surface

“…A limitation of the reported method is that N-glycans must be released by PNGaseF from denatured proteins to be a substrate for galactosidase. In 2018, Butler et al reported that a galactosidase from Bacteroides fragilis could remove terminal galactose directly from naïve antibody thus could be applied for high yields of single glycoform antibodies[20, 21]. However, all the previous studies on galactosidase activity were conducted in a non-cell system.…”