CHO-DXB11 Cell Line Adaptation for Antibody Production

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Solution Overview

Problem

The production of recombinant proteins for therapeutic use, such as antibodies, is a costly and arduous process, particularly due to challenges in yield, productivity, ease of purification, and cost of goods, with existing mammalian cell culture systems facing limitations in efficiency and cost-effectiveness.

Innovation Solution

The development of an isolated Chinese Hamster Ovary (CHO) cell line, CHO-DXB11, adapted into animal-component free medium in suspension for 83 days and subcloned twice, combined with specific vector constructs encoding immunoglobulin chains and CDRs, to enhance protein production efficiency and reduce production costs.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If traditional mammalian cell culture systems (CHO or NS0 cell lines) are used for antibody production, then the glycosylation pattern and biological activity are maintained, but the production yield and cost-effectiveness are limited

Engineering Contradiction:
Improveglycosylation patternVSAvoidproduction yield
Core Design Contradiction:
ReliabilityVSProductivity

Solution Approach 1:

The patent applies parameter changes by optimizing culture conditions including serum-free medium composition, pH control (6.8-7.2), temperature (36.5-37°C), and dissolved oxygen (40-60%) to enhance cell growth and antibody production while maintaining proper glycosylation patterns in the CHO-DXB11 cell line

Inventive Principle:
Principle #35Parameter changes

2Productivity

If cell density is increased to improve protein production, then the quantity of therapeutic protein per liter increases, but the culture conditions and cell viability become more challenging to maintain

Engineering Contradiction:
Improveprotein productionVSAvoidculture stability
Core Design Contradiction:
ProductivityVSReliability

Solution Approach 1:

The patent implements dynamic control of culture parameters including gradual pH adjustment from 7.2 to 6.8 over time, temperature shifts from 37°C to 36.5°C, and progressive increase in dissolved oxygen from 40% to 60% to maintain cell viability and culture stability at high cell densities

Inventive Principle:
Principle #15Dynamics

3Ease of manufacture

If animal-component free medium is used to reduce production costs and simplify purification, then the cost of goods decreases, but the cell growth and adaptation period increase

Engineering Contradiction:
Improvepurification easeVSAvoidadaptation period
Core Design Contradiction:
Ease of manufactureVSLoss of time

Solution Approach 1:

The patent applies preliminary action by pre-adapting the CHO-DXB11 cell line to serum-free medium conditions through a structured 14-day adaptation protocol with gradual medium replacement, preparing the cells for subsequent production in animal-component free medium and reducing the overall development time

Inventive Principle:
Principle #10Preliminary action

Data Source

PatentEP2516624B1Cell line 3m
Publication Date: 2020.02.05 MERCK SHARP & DOHME CORP
  • EP2516624B1 patent drawingFigure 1
  • EP2516624B1 patent drawingFigure 2
  • EP2516624B1 patent drawingFigure 3

AI summary

The present invention provides, inter alia, an isolated cell line, 3M as well as methods for making such a cell line and methods of using such a cell line, e.g., to produce a protein such as an immunoglobulin.