@misc{10481/92044,
year = {2024},
month = {2},
url = {https://hdl.handle.net/10481/92044},
abstract = {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.},
organization = {UKRI Engineering and Physical
Sciences Research Council (EP/N509486/1)},
organization = {UKRI Engineering and Physical Sciences Research Council
(EP/K038648/1, EP/H04986X/1 and EP/K038648/1)},
organization = {UK Research and Innovation ‘Global Challenges
Research Fund’ BB/P02789X/1)},
organization = {UK Biotechnology
and Biological Sciences Research Council},
publisher = {Springer Nature},
title = {Immobilized enzyme cascade for targeted glycosylation},
doi = {10.1038/s41589-023-01539-4},
author = {Makrydaki, Elli and Donini, Roberto and Krueger, Anja and Royle, Kate and Moya-Ramírez, Ignacio and Kuntz, Douglas A. and Rose, David R. and Haslam, Stuart M. and Polizzi, Karen M. and Kontoravdi, Cleo},
}