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

Methods for genetic engineering kluyveromyces host cells

A technology of Kluyveromyces and host cells, applied in the field of genetic engineering Kluyveromyces host cells, which can solve the surge in demand for monoclonal antibodies, long timeline for new antibodies, and CHO cell line productivity is unlikely to be significantly improved And other issues

Pending Publication Date: 2020-06-30
AMYRIS INC
View PDF9 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Furthermore, after decades of refinement, CHO cell line productivity is unlikely to increase significantly, and with the introduction of therapies for broader and chronic conditions (e.g., Alzheimer's disease), the demand for monoclonal antibodies is expected to increase. will surge
Therefore, there are two problems with CHO cells: a long timeline for producing new antibodies, and insufficient capacity to meet future needs

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
  • Methods for genetic engineering kluyveromyces host cells
  • Methods for genetic engineering kluyveromyces host cells
  • Methods for genetic engineering kluyveromyces host cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0118] In the present invention, secretion of two full-length antibodies at high titers (≥19 μg / mL) in Kluyveromyces marxe: Herceptin (trastuzumab) and MabThera (rituximab) has been achieved . High titers were achieved using codon optimization of the amino acid sequences of the heavy chain, light chain and secretion tag according to K. marx preferred codons. The DNA expression constructs were cloned as convergent split cassettes at the same locus under a strong constitutive native glycolytic promoter. The copy number of the cassette (2 / 3 copies seemed optimal) was variable. Using 4% glucose (from sugarcane source), cell recycle treatment significantly improved titers. In addition, a K. marikus production strain with three copy numbers of Herceptin was run in a 0.5L fermenter and high titers of full-length antibodies were obtained (estimated titer ≥ 30 μg / mL). Strains were constructed using CRISPR tools with deletion of the NHEJ pathway. The copy number of gene integration ...

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

No PUM Login to View More

Abstract

The invention provides methods for genetically engineering Kluyveromyces. The methods can be used to genetically engineer Kluyveromyces to produce and secrete full-length antibodies or antibody fragments. The invention also provides methods for production and secretion of antibodies.

Description

[0001] field of invention [0002] The methods and compositions provided herein relate generally to the fields of molecular biology and genetic engineering. Background technique [0003] Genetic engineering techniques for introducing targeted modifications into the genome of host cells can be used in a variety of fields. Fundamentally, determining how genotype affects phenotype depends on the ability to introduce targeted insertions or deletions that impair or abolish native gene function. In the field of synthetic biology, making genetically modified microorganisms capable of producing a compound or protein of interest requires the insertion of a custom DNA sequence into the host cell's chromosome; industrial-scale production often requires the introduction of multiple genes into the host cell genome. [0004] For certain host cells, especially for conventional yeast cells (eg, Saccharomyces cerevisiae), genetic tools are well developed for targeted genomic gene deletion and...

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(China)
IPC IPC(8): C12N15/90
CPCC12N15/905C07K16/00C12N15/815C12N2310/20C12N15/102C07K16/2851C07K16/2887C07K16/2893C07K16/32C07K2317/14C12N9/22C12N15/113C12N2800/80
Inventor Y·茨嘎也
Owner AMYRIS INC
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