Method for in Vivo Production of Deglycosylated Recombinant Proteins Used as Substrate for Downstream Protein Glycoremodeling
AssigneeCaliber Biotherapeutics, LLC
InventorsSylvain Marcel, Lindsay Bennett
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Abstract
The present invention includes compositions and methods of reducing the glycosylation of proteins comprising: obtaining a cell that expresses one or more proteins that comprise one or more glycosylation sites and are glycosylated; expressing in the cell one or more glycosidases that cleaves one or more glycosyl groups from the one or more proteins; and isolating the one or more proteins with reduced glycosylation from the cell.
Claims35
A method of reducing the glycosylation of proteins comprising:
- obtaining a cell that expresses one or more proteins that comprise one or more glycosylation sites and are glycosylated;
- expressing in the cell one or more glycosidases that cleave one or more glycosyl groups from the one or more proteins to reduce the glycosylation of the protein, wherein the glycosidase;
- and
- isolating the one or more proteins with reduced glycosylation from the cell.
A method of reducing the glycosylation of proteins comprising:
- co-expressing one or more proteins that comprise one or more glycosylation sites and are glycosylated in a cell with one or more glycosidases, wherein the one or more glycosidases act to reduce or eliminate the glycosylation of the one or more proteins in the cell;
- and
- purifying the one or more proteins from the cell, wherein the glycosidase acts on glycosylation on the one or more proteins.
A method of reducing the glycosylation of proteins comprising:
- co-expressing one or more proteins that comprise one or more glycosylation sites and are glycosylated and one or more glycosidases that cleaves one or more glycosylations from the one or more proteins in a cell;
- and
- purifying the one or more proteins from the cell, wherein the glycosidase acts on glycosylation on the one or more proteins;
- and
- reglycosylating the one or more proteins using a mutant glycosidase.
A glycoprotein made by a method comprising:
- obtaining a cell that expresses one or more proteins that comprise one or more glycosylation sites and are glycosylated;
- expressing in the cell one or more glycosidases that cleaves one or more glycosyl groups from the one or more proteins to reduce the glycosylation of the protein;
- and
- isolating the one or more proteins from the cell, wherein the glycosidase acts on glycosylation on the one or more proteins in the cell to reduce the glycosylation of the one or more proteins.
A glycoprotein made by a method comprising:
- expressing a protein in a plant cell that is recombinantly modified to expresses a glycosidase;
- isolating the protein expressed and deglycosylated in the cell;
- re-glycosylating the protein in the presence of one or more saccharides in at least one of in vitro or in cellulo;
- and
- isolating the glycosylated protein.
Dependencies are inferred from the claim text.