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In Vitro Sialylation of Recombinant Alpha-1-Antitrypsin using α2,6 Sialyltransferase from Photobacterium Damselae Produces Di-sialyl Galactose Motifs in N-Glycans

preprint
submitted on 07.06.2019 and posted on 07.06.2019 by Edward Pallister, Matthew Choo, jien nee tai, Dawn leong, Wen-Qin Tang, Say-Kong Ng, Kun Huang, Andrea Marchesi, Peter Both, Chris Gray, Pauline Rudd, Sabine Flitsch, Terry Nguyen-Khuong
Sialic acids are cell surface sugars present in many animal glycoproteins and are of particular interest in biopharmaceuticals, where lack of sialylation can reduce bioactivity. Here we describe how α-2,6-sialyltransferase from Photobacterium Damselae can be used to markedly increase sialylation of CHO produced alpha-1-antitrypsin. Detailed analysis of the sialylation products showed that in addition to the expected α-2,6-sialylation of galactose, a second di-sialyl galactose motif was produced, which had never been recorded on a mammalian glycoprotein. The influence of this unique di-sialylation on the in vitro activity of alpha-1-antitrypsin was studied and a toolkit of mass spectrometry methods to identify this new di-sialyl galactose motif in complex mixtures was developed

Funding

A*STAR GlycoSing

A*STAR HighGlycoArt

A*STAR ARAP

History

Email Address of Submitting Author

edward.pallister@gmail.com

Institution

A*STAR Bio-processing Technology Institute

Country

Singapore

ORCID For Submitting Author

0000-0002-9681-7327

Declaration of Conflict of Interest

None

Version Notes

V1 intial draft

Exports