Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Mammalian cell culture process

a cell culture process and mammalian technology, applied in the field of mammalian cell culture process, can solve the problems of increasing the sialic acid content of mature proteins, little guidance in selecting appropriate concentrations of additives, and not addressing the effect of additives on protein glycosylation

Inactive Publication Date: 2004-12-23
ETCHEVERRY TINA +1
View PDF9 Cites 7 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

"This patent relates to processes for controlling the sialic acid content of glycoproteins produced in mammalian cell culture. The patent discusses how to increase or decrease the amount of sialic acid in glycoproteins to improve their function and make them suitable for use as therapeutics. The patent also describes methods for producing tumor necrosis factor receptor-immunoglobulin chimeras and novel TNFR1-IgG preparations for the diagnosis and treatment of inflammatory and immune disorders. The patent highlights the importance of maintaining the sialic acid content of glycoproteins for their function and physical, chemical, and immunogenic properties. The patent also discusses the impact of various factors such as growth mode, fetal bovine serum, and pH on glycosylation and provides guidance on selecting appropriate concentrations of alkanoic acids for enhanced protein production."

Problems solved by technology

The publication, however, provides little guidance in selecting appropriate concentrations of the additive and further does not address the effect of the additive on protein glycosylation.
However, the previous studies have not studied the effect of various process parameters on the sialic acid content of the mature protein, a factor in glycoprotein production that is tantamount to clinical success.
Conversely, a decrease in cell specific productivity results in an increase in sialic acid content in the mature protein.
The lower concentration provides for enhanced transcription, but maintains lower cell specific productivity while maintaining the viability of the host cell culture.

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
  • Mammalian cell culture process
  • Mammalian cell culture process

Examples

Experimental program
Comparison scheme
Effect test

examples

[0121] The biological effects of TNF-alpha and TNF-beta are mediated through specific receptors (Dembic et al., (1990) Cytokines, 2:231). Molecular cloning has demonstrated the existence of two distinct types of TNF receptors (TNFR) with apparent molecular masses of 55kD (type 1) (Schall et al., (1990) Cell 61:361) and 75 kD (type 2) (Smith et al., (1990) Science 248:1019), each of which naturally binds to both TNF alpha and TNF beta (Loetscher et al., (1990) Cell, 61:351; Shall et al. (1990) Cell, 61:361; Kohno et al., (1990) Proc. Natl. Acad. Sci. USA 87:8331). The extracellular portions of both receptors are found naturally as soluble TNF binding proteins (Kohno et al., supra). TNF agonists have been created which block the deleterious effect of TNF in various immune and inflammatory events (Peppel et al., (1991) J. Exp. Med., 174:1483-1489; Ulich (1993) Am. J. Path., 142:1335-1338; Howard, O. M. Z., (1993) Proc. Natl. Acad. Sci. USA 90:2335-2339; Wooley, P. H., (1993) J. Immunol...

example ii

[0144] Plasma pharmacokinetic half-lives and / or clearance rates of different preparations were determined. The preparations containing a higher sialic acid content in general exhibited increased plasma half-life and / or lower total clearance rates compared to preparations containing a decreased sialic acid content.

[0145] Methods

[0146] Seventeen male Sprague-Dawley-derived rats weighing 272-315 g were randomly assigned to one of three TNFR1-IgG.sub.1 fusion protein treatment groups (N=5 or 6 per group). The animals were injected intravenously with a nominal dose of 5 mg / kg of test material via a femoral vein cannula. The test materials chosen included two TNFR1-IgG.sub.1 preparations from a process in which the butyrate concentration during the production phase was 6 mM and the osmolality during the production phase was maintained at about 400 mOsm, Process I, and a third TNFR1-IgG.sub.1 preparation from a process in which the butyrate concentration was 12 mM and the osmolality during...

example iii

Monosaccaride Composition of TNFR1-IgG.sub.1

[0151] Determination of oligosaccharide carbohydrate composition and structure of TNFR1-IgG.sub.1, prepared as described in Example 1 showed that the sialic acid content of the various process version varied. Rapid plasma clearance was associated with high exposed GlcNAc, lower sialic acid on the oligosaccharide chains and (by inference) accessibility of the protein to mannose or galactose receptors. Slow plasma clearance was associated with more terminal sialic acid residues.

[0152] A. Sources of Test Material

[0153] TNFR1-IgG.sub.1 was produced according to the methods outlined as described in Example 1 above. Process I material was obtained from cell culture using 6 mM butyrate and an osmolality of about 400 mOsm during the production phase. Process material II was obtained from cell culture using 12 mM butyrate and an osmolality of about 500 mOsm during the production phase.

[0154] B. Methods

[0155] Release of intact neutral and amino-suga...

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
molar ratioaaaaaaaaaa
molar ratioaaaaaaaaaa
temperatureaaaaaaaaaa
Login to View More

Abstract

The present invention relates to novel process for the preparation of glycoproteins by mammalian cell culture wherein the sialic acid content of the glycoprotein produced is controlled over a broad range of values by manipulating the cell culture environment. The invention provides for processes in which the sialic acid content of the glycoprotein is modified by changes in cell culture parameters which affect cell specific productivity. Preferred embodiments of the invention include cell culture processes in the osmolality of the cell culture is controlled as well as the concentration of a transcription enhancer during the production phase of the cell culture. The invention further provides for novel preparations of soluble type 1 tumor necrosis factor immunoglobulin G1 and their uses in the treatment of inflammatory or immune related disorders.

Description

[0001] This invention relates to processes for controlling the sialic acid content of glycoproteins produced in mammalian cell culture. The invention provides processes for increasing and decreasing the sialic acid content of glycoproteins produced by mammalian cell culture. The invention further relates to processes for producing tumor necrosis factor receptor (TNFR)-immunoglobulin (Ig) chimeras as well as novel TNFR1-IgG.sub.1 preparations and their uses in the diagnosis and treatment of various inflammatory and immune disorders.DESCRIPTION OF RELATED ART[0002] Differences in glycosylation patterns of recombinantly produced glycoproteins have recently been the topic of much attention in the scientific community as recombinant proteins produced as probable prophylactics and therapeutics approach the clinic. The oligosaccharide side chains of the glycoproteins affect the protein's function (Wittwer A., and Howard, S. C. (1990) Biochem. 29:4175-4180) and the intramolecular interactio...

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
Patent Type & Authority Applications(United States)
IPC IPC(8): C07K14/715
CPCC07K14/7151C07K2319/00C07K2319/30
Inventor ETCHEVERRY, TINARYLL, THOMAS
Owner ETCHEVERRY TINA
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com