US 11,078,511 B2Granted

Aqueous composition

AssigneeRoche Diagnostics Operations, Inc.

InventorsHarald Sobek, Michael Greif, Marco Thomann, Sebastian Malik

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Abstract

The present disclosure is directed to the properties of certain glycosyltransferase variants having N-terminal truncation deletions or internal deletions. Any of the mutants disclosed in here exhibit alpha-2,6-sialyltransferase enzymatic activity in the presence of CMP-activated sialic acid as co-substrate, and in the presence of a suitable acceptor site. A fundamental finding documented in the present disclosure is that these enzymes are not only capable of catalyzing transfer of a sialidyl moiety but they are also capable of catalyzing hydrolytic cleavage of terminally bound sialic acid from a glycan.

Claims1

Dependent claims

An aqueous composition comprising:

(a) a soluble human β-galactoside-α-2,6-sialyltransferase I comprising the amino acid motif from position 90 to position 108 in SEQ ID NO:1;
(b) a cytidine-5′-monophospho-N-acetylneuraminic acid;
(c) a glycosylated target molecule selected from the group consisting of a glycoprotein and a glycolipid, the target molecule comprising one or more antenna(e), at least one antenna having as a terminal structure a β-D-galactosyl-1,4-N-acetyl-β-D-glucosamine moiety with a hydroxyl group at the C6 position in the galactosyl residue;
(d) an aqueous solution permitting glycosyltransferase enzymatic activity comprising a buffer salt;
wherein the aqueous composition further comprises an enzyme selected from the group consisting of an alkaline phosphatase, an acid phosphatase, and a 5′ nucleotidase capable of hydrolyzing the phosphoester bond in 5′-cytidine-monophosphate under conditions permitting glycosyltransferase enzymatic activity.

2 more elements

Dependencies are inferred from the claim text.

Facts

Application: 15/629,040Filed 21 Jun 2017 · continuation
Priority: 21 Dec 2015
Published: US 2017/0298405 A119 Oct 2017
Granted: 3 Aug 2021
Term ends: Not estimatedThe source records don’t support an estimate.

Classification11 codes

C12P 21/005
Preparation of peptides or proteins: glycopeptides, glycoproteins
C07K 16/00
Immunoglobulins [ig]
C12N 9/1081
Glycosyltransferases: transferring other glycosyl groups
C12P 19/44
Preparation of compounds containing saccharide radicals: preparation of O-glycosides, e.g. glucosides

Family

PCT/EP2015/080741Parent of this continuation · filed 21 Dec 2015
US 2017/0298405 A1Pre-grant publication · published 19 Oct 2017
US 11,078,511 B2This patent

From the US filing history. Foreign family members aren’t listed.

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