Deglycosylated enzymes for conjugates
AssigneeRoche Diagnostics Operations, Inc.
InventorsWolfgang Hoesel, Werner Hoelke, Beata Kanne, Helmut Lenz, Rainer Mueller
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Abstract
By deglycosylating an enzyme derived from a eukaryotic organism prior to forming a conjugate, i.e. prior to attaching the enzyme to a molecule capable of binding to a target molecule, the sensitivity of an assay for detecting the presence or determining the quantity of a target molecule, in which the conjugate is used as a labelled component, can be increased. The present invention provides conjugates, methods for providing the same, use of such conjugates as well as kits containing the conjugates.
Claims19
A method to produce a conjugate of an alkaline phosphatase enzyme and a molecule which binds to a target molecule comprising the steps of:
- (a) providing a glycosylated alkaline phosphatase enzyme obtained by expression in a transformed yeast and isolated therefrom,
- (b) deglycosylating the glycosylated enzyme of step (a),
- (c) isolating the deglycosylated enzyme, and
- (d) attaching the deglycosylated enzyme of step (c) to the molecule capable of binding to a target molecule.
An isolated and partially deglycosylated alkaline phosphatase enzyme of mammalian origin, whereby the enzyme is free of mammalian proteins, obtainable by the method comprising the steps of:
- (a) providing a glycosylated alkaline phosphatase enzyme obtained by expression in a transformed yeast and isolated therefrom, whereby the carbohydrate portion of the glycosylated enzyme contains multiple mannose subunits,
- (b) partially deglycosylating the glycosylated enzyme of step (a), and
- (c) isolating the partially deglycosylated enzyme.
Dependencies are inferred from the claim text.