Rapid preparation of a glycan oxazoline and a homogeneously glycosylated antibody with an enzyme-immobilized monolithic column

Chemo-enzymatic glycan engineering is considered to be one of the most promising strategies to enhance efficiency in pharmaceutical research. However, it is assumed that this technology has limited industrial application for the production of biological therapeutics because of the high cost of the process. In this study, we developed a scheme for rapidly preparing a glycan oxazoline and a homogeneously glycosylated antibody. The enzyme-immobilized monolith and the flow chemistry-based approach enabled a glycan oxazoline and a homogeneously glycosylated antibody to be obtained at the gram scale from starting materials (sialylglycopeptide and heterogeneously glycosylated protein) within 2.5 h. This cost-effective scheme for obtaining a large amount of glycan donors and homogeneously glycosylated proteins in a short time will be helpful to implement glycan engineering technology for industrial purposes such as pharmaceutical production. [Display omitted] •A rapid glycan engineering scheme based on immobilized enzyme and flow chemistry.•ENGases were immobilized on monolith via a histidine tag and a metal ion.•A glycan oxazoline and a homogeneously glycosylated antibody were obtained.•Gram scale preparation was achieved within 2.5 h from starting materials.