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

a technology of recombinant glycoproteins and insect cells, which is applied in the field of molecular biology and the production of glycoproteins lacking fucosylation, can solve the problems of no reported efforts to block the fucosylation of recombinant glycoproteins in any insect-based system, and pre-existing human antibodies can give false positives, etc., to achieve the effect of stabilizing the inhibition of fucosylation

Inactive Publication Date: 2015-04-02
UNIVERSITY OF WYOMING
View PDF2 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0014]In a further embodiment, the invention provides a method for production of a non-fucosylated protein in insect larvae. An exemplary method entails providing insect larvae and introducing therein a baculovirus comprising at least one nucleic acid molecule encoding a codon optimized enzyme GDP-4-dehydro-6-deoxy-D-mannose reductase (RMD) operably driven by an immediate early expression control sequence for expression immediately after infection or an inducible promoter, thereby stabilizing inhibition of fucosylation, and at least one additional nucleic ...

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...

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Compositions and methods for reducing fucosylation of glycoproteins in insect cells and methods of use thereof for production of recombinant glycoproteins
  • Compositions and methods for reducing fucosylation of glycoproteins in insect cells and methods of use thereof for production of recombinant glycoproteins
  • Compositions and methods for reducing fucosylation of glycoproteins in insect cells and methods of use thereof for production of recombinant glycoproteins

Examples

Experimental program
Comparison scheme
Effect test

example i

A Novel Baculovirus Vector for the Production of Non-Fucosylated Recombinant Glycoproteins in Insect Cells

[0066]Analysis of Core α1,3-Fucosylation in Three Insect Cell Lines

[0067]High Five™ cells, derived from Trichoplusia ni, but not Sf9 cells, derived from Spodoptera frugiperda, produce core α1,3-fucosylated glycoproteins (Rudd et al. 2000; Seismann et al. 2010; Blank et al. 2011; Palmberger et al. 2011). Another Trichoplusia ni cell line used as a host for baculovirus expression vectors is Tni PRO™ (Kwon et al. 2009; Bourhis et al. 2010; Bongiovanni et al. 2012; He et al. 2013; Merchant et al. 2013), but its capacity for core α1,3-fucosylation has not been reported. Thus, we analyzed intracellular extracts of uninfected Tni PRO™ cells by western blotting with anti-horseradish peroxidase (HRP), which detects core α1,3-linked fucosylation, using extracts from Sf9 and High Five™ cells as negative and positive controls. Coomassie brilliant blue staining showed that approximately equa...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
Therapeuticaaaaaaaaaa
Login to view more

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...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12N9/04C12P21/00
CPCC12N9/0006C12Y101/01281C12P21/00A61K38/00C07K16/2887C07K2317/14C07K2317/24C07K2317/41C12P21/005
Inventor MABASHI-ASAZUMA, HIDEAKIJARVIS, DONALD L.
Owner UNIVERSITY OF WYOMING
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap