ABC50 Overexpression for Recombinant Protein Yield

Resolve Bottlenecks,
Find Innovative SolutionsGenerate Solutions

Solution Overview

Problem

Current methods for producing recombinant proteins, particularly monoclonal antibodies, face challenges in optimizing protein production levels in mammalian cells, leading to inefficiencies in therapeutic and diagnostic applications.

Innovation Solution

The method involves increasing the expression or activity of the ABC50 protein or its fragments with eIF2 binding activity, either by introducing a heterologous ABC50 polynucleotide linked to a promoter or using Econazole to enhance protein synthesis and production in cells, thereby increasing the yield of specific proteins like antibodies.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If conventional methods are used to produce recombinant proteins in mammalian cells, then protein production can be achieved, but production levels are insufficient and efficiency is low

Engineering Contradiction:

Improveprotein production levelVSAvoidproduction efficiency

Core Design Contradiction:

ProductivityVSEase of manufacture

Solution Approach 1:

The patent changes the biochemical parameters of the cell by overexpressing ABC50 protein, which modifies translation initiation efficiency. This parameter change enables significantly higher protein production levels (up to several hundred percent increase) while maintaining production efficiency, directly resolving the contradiction between productivity and ease of manufacture.

Inventive Principle:

Principle #35Parameter changes

2Productivity

If ABC50 expression is increased to boost protein production, then protein yield increases, but cell sensitivity to ER stress agents changes

Engineering Contradiction:

Improveprotein yieldVSAvoidcell sensitivity to ER stress

Core Design Contradiction:

ProductivityVSObject-affected harmful factors

Solution Approach 1:

The patent converts the harmful effect of ER stress agent sensitivity into a beneficial selection mechanism. By overexpressing ABC50, cells become more sensitive to ER stress agents like Econazole, which can be used as a selective pressure to maintain the high-producing cell line. The increased sensitivity, while seemingly harmful, becomes a useful tool for ensuring consistent high protein production.

Inventive Principle:

Principle #22Blessing in disguise (Convert harm into benefit)

Applied Scientific Principles

This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.

Function Achieved in This Case

This approach results in significant increases in protein production, up to several hundred percent, improving the efficiency of recombinant protein production and enhancing the sensitivity of cells to ER stress agents.

Implementation Method 1

ABC50 stimulates formation of complexes between eIF2, GTP and Met-tRNA, implicating it in translation initiation and control

Methodology Applied

Scientific EffectTranslation initiation:

Implementation Method 2

Econazole (Ec) is an imidazole antifungal that also induces endoplasmic reticulum (ER) stress by promoting ER Ca2+ depletion

Methodology Applied

Scientific EffectER stress:

Data Source

PatentUS8691529B2Increasing protein production by increasing ABC50 expression or activity

Publication Date: 2014.04.08 UNIV HEALTH NETWORK

AI summary

The disclosure provides methods and materials for increasing the expression of a protein of interest such as an antibody by a cell ABC50 expression or activity is increased which increases expression of the protein or antibody of interest. The disclosure also provides methods and materials for increasing the sensitivity of a cell to an endoplasmic reticulum stress agent such as Econazole by decreasing the level of ABC50.