Compositions and methods for reducing fucosylation of glycoproteins in insect cells and methods of use thereof for production of recombinant glycoproteins

Abstract

Compositions for reducing fucosylation of glycoproteins in insect cells are provided. Also disclosed are methods of use of such compositions for the production of recombinant humanized proteins.

Description

PRIORITY CLAIM[0001]This application claims priority to U.S. Provisional Application Number 61 / 885,294 filed Oct. 1, 2013. This application is incorporated herein by reference as though set forth in full.STATEMENT OF RIGHTS TO INVENTIONS MADE UNDER FEDERALLY SPONSORED RESEARCH[0002]Pursuant to 35 U.S.C. §202(c) it is acknowledged that the U.S. Government has rights in the invention described, which was made with funds from the National Institutes of Health, R01GM49734.FIELD OF THE INVENTION[0003]This invention relates to the fields of molecular biology and production of glycoproteins lacking fucosylation. More specifically, the invention provides compositions and methods for expressing a GDP-4-dehydro-6-deoxy-D-mannose reductase enzyme encoded by a recombinant baculovirus vector that blocks the production of GDP-L-fucose and generates a molecule lacking or having a reduced amount of fucose.BACKGROUND OF THE INVENTION[0004]Numerous publications and patent documents, including both pu...

AI Technical Summary

Benefits of technology

The patent is about a method for making non-fucosylated proteins in insect larvae. The method involves introducing a baculovirus carrying genes for a protective enzyme and a protein of interest into the insect. The enzyme prevents the production of a compound that adds fucose to the protein. This results in a non-fucosylated protein that can be isolated. The technical effect of this method is that it provides a way to produce proteins with a specific modification that is important for their function in the animal or plant they are intended for use in.

Problems solved by technology

In addition, pre-existing human antibodies against the immunogenic sugar epitope can give false positive results in diagnostic tests that use α1,3-fucosylated recombinant glycoproteins produced in the baculovirus-insect cell system (Hancock et al.

To date, however, there have been no reported efforts to block recombinant glycoprotein fucosylation in any insect-based system, including the baculovirus-insect cell system, despite the arguably more serious problem of immunogenicity associated with core α1,3-fucosylation mediated by some insect and insect cell types.

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