Citation

BibTex format

@article{Makrydaki:2024:10.1038/s41589-023-01539-4,
author = {Makrydaki, E and Donini, R and Krueger, A and Royle, K and Moya, Ramirez I and Kuntz, DA and Rose, DR and Haslam, SM and Polizzi, KM and Kontoravdi, C},
doi = {10.1038/s41589-023-01539-4},
journal = {Nature Chemical Biology},
title = {Immobilized enzyme cascade for targeted glycosylation},
url = {http://dx.doi.org/10.1038/s41589-023-01539-4},
year = {2024}
}

RIS format (EndNote, RefMan)

TY  - JOUR
AB - Glycosylation is a critical post-translational protein modification that affects folding, half-life and functionality. Glycosylation is a non-templated and heterogeneous process because of the promiscuity of the enzymes involved. We describe a platform for sequential glycosylation reactions for tailored sugar structures (SUGAR-TARGET) that allows bespoke, controlled N-linked glycosylation in vitro enabled by immobilized enzymes produced with a one-step immobilization/purification method. We reconstruct a reaction cascade mimicking a glycosylation pathway where promiscuity naturally exists to humanize a range of proteins derived from different cellular systems, yielding near-homogeneous glycoforms. Immobilized β-1,4-galactosyltransferase is used to enhance the galactosylation profile of three IgGs, yielding 80.2-96.3% terminal galactosylation. Enzyme recycling is demonstrated for a reaction time greater than 80 h. The platform is easy to implement, modular and reusable and can therefore produce homogeneous glycan structures derived from various hosts for functional and clinical evaluation.
AU - Makrydaki,E
AU - Donini,R
AU - Krueger,A
AU - Royle,K
AU - Moya,Ramirez I
AU - Kuntz,DA
AU - Rose,DR
AU - Haslam,SM
AU - Polizzi,KM
AU - Kontoravdi,C
DO - 10.1038/s41589-023-01539-4
PY - 2024///
SN - 1552-4450
TI - Immobilized enzyme cascade for targeted glycosylation
T2 - Nature Chemical Biology
UR - http://dx.doi.org/10.1038/s41589-023-01539-4
UR - https://www.ncbi.nlm.nih.gov/pubmed/38321209
UR - http://hdl.handle.net/10044/1/109176
ER -