Lectin-based glycan assay
AssigneeGalab Technologies GmbH
InventorsJürgen Kuballa, Kirstin Umnus, Simone Cartellieri
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Abstract
The invention relates to the use of deglycosylated detectors, especially secondary antibodies, for the determination of sugar structures of proteins, especially recombinant proteins. It further relates to the use of deglycosylated enzymes in the determination of sugar structures with the aid of an enzyme-substrate reaction; the invention further relates to a method of determining the sugar structures of proteins, a sugar determination kit and the use of said kit for the determination of sugar structures, especially of recombinant therapeutic proteins, preferably immunoglobulins.
Claims13
A method for the determination of the sugar structures of recombinant proteins, wherein the method comprises the following steps:
- a) providing a recombinant protein to be investigated, which is a recombinant therapeutic glycoprotein, said protein being present in immobilized form, through a deglycosylated Pectin, wherein the lectin is present immobilized on a support;
- b) incubating said recombinant protein with a labelled binding structure including at least one of, a secondary antibody or a protein A or fragment thereof, which is deglycosylated;
- and
- c) determining the binding between said recombinant protein and the binding structure wherein the sugar structure of said recombinant protein can be derived from said lectin.
A method for the determination of the sugar structures of immunoglobulins, wherein the method comprises the following steps:
- a) providing the immunoglobulin to be investigated, said immunoglobulin being present in immobilized form, through a deglycosylated lectin, wherein the lectin is present immobilized on a microtiter plate,
- b) incubating said immunoglobulin with a labeled binding structure including at least one of, a secondary antibody or a protein A or fragment thereof, which is deglycosylated;
- and
- c) determining the binding between the immunoglobulin and the binding structure wherein the sugar structure of said recombinant protein can be derived from said lectin.
A method for the determination of the sugar structures of recombinant proteins, wherein the method comprises the following steps:
- a) providing the recombinant protein to be investigated, which is a recombinant therapeutic glycoprotein, said protein being present in immobilized form, through a deglycosylated lectin, wherein the lectin is present immobilized on a microtiter plate;
- said immobilization of said recombinant protein by means of lectin occurs when said recombinant protein comprises a sugar structure that is recognized and bound by said lectin;
- b) incubating said recombinant protein with a labelled binding structure including at least one of, a secondary antibody or a protein A or fragment thereof, which is deglycosylated;
- and
- c) determining the binding between the said recombinant protein and the binding structure wherein the sugar structure of said recombinant protein can be derived from said lectin.
Dependencies are inferred from the claim text.