Momenta Pharmaceuticals, Inc.

Ian Christopher Parsons

Carlos J Bosques

Nur Sibel Gunay

Ting Zheng

Jonathan C Lansing

Brian Edward Collins

Dorota A Bulik

Lakshmanan Thiruneelakantapillai

Daniela Beccati

Xiangping Zhu

Scott Bailey

Zachary Shriver

Ganesh Venkataraman

Nathaniel J Washburn

Rajeev Chillakuru

Daniela Becatti

Zachary Schriver

A multi-dimensional chromatographic method for the separation of N-glycans. The method comprises providing a glycan preparation that includes at least one negatively charged N-glycan. The glycan preparation is then separated by anion-exchange chromatography and at least one secondary chromatographic technique.

Multi-dimensional chromatographic methods for separating N-glycans

The present disclosure provides a method for determining the relative conformations of a protein provided in different protein preparations, comprising steps of: (i) obtaining a first 2D NOESY NMR spectrum of a first protein preparation; (ii) obtaining a second 2D NOESY NMR spectrum of a second protein preparation; and (iii); determining whether a protein has a different conformation in the first and second protein preparations by comparing one or more cross-peaks in the first 2D NOESY NMR spectrum with one or more corresponding cross-peaks in the second 2D NOESY NMR spectrum.

Comparative analysis of protein conformations by using 2D NOESY NMR spectra

The present disclosure provides methods for analyzing structure and/or composition of N-glycans. Such methods often involve digestion of N-glycans with multiple exoglycosidases. In some embodiments, N-glycans are digested with multiple exoglycosidases simultaneously. In some embodiments, N-glycans are digested with multiple exoglycosidases sequentially. In some embodiments, methods in accordance with the present disclosure involve comparison of cleavage products of N-glycans that have been digested with multiple exoglycosidases simultaneously to N-glycans that have been digested with multiple exoglycosidases sequentially.

Characterization of N-glycans using exoglycosidases

The present disclosure provides nuclear magnetic resonance (NMR) methods for characterizing mixtures of N-linked glycans. Without limitation, methods of the present disclosure may be useful in characterizing monosaccharide composition, branching, fucosylation, sulfation, phosphorylation, sialylation linkages, presence of impurities and/or efficiency of a labeling procedure (e.g., labeling with a fluorophore such as 2-AB). In certain embodiments, the methods can be used quantitatively. In certain embodiments, the methods can be combined with enzymatic digestion to further characterize glycan mixtures.

Characterization of N-glycan mixtures by nuclear magnetic resonance

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The present disclosure provides methods for assessing the glycosylation of a target glycoprotein produced by a cell through analysis of cell-surface glycans on the cell. The present disclosure therefore teaches that glycosylation of cell surface proteins can serve as a proxy for glycosylation of other proteins.

Methods related to cell surface glycosylation

The present disclosure provides, among other things, methods for the identification of sulfated glycans in a mixture of glycans.

MS methods to evaluate glycans

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Belmont, MA, United States of America

Ian Christopher Parsons