Egg White Acidification for Bulk Protein Isolation

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Solution Overview

Problem

Traditional methods for isolating exogenous proteins from egg white, such as those from transgenic chickens, are not practical for large-scale production due to high costs, labor, and time requirements, especially when dealing with industrial volumes exceeding 10 liters.

Innovation Solution

Adding a small amount of acidic buffer (0.5-5 wt% per kilogram) to egg white to adjust the pH to 5-6.5, reducing the volume of egg white subject to downstream processes, allowing for bulk chromatographic isolation without significant dilution, and facilitating the separation of recombinant proteins.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Manufacturing precision

If traditional immunoaffinity procedures are used to isolate exogenous proteins from egg white, then purification can be achieved, but the process becomes impractical for large-scale production due to high costs, labor, and time requirements

Engineering Contradiction:
Improvepurification qualityVSAvoidproduction scale
Core Design Contradiction:
Manufacturing precisionVSProductivity

Solution Approach 1:

The patent changes the pH parameter of egg white from its natural state to an acidic range (pH 2-4) through addition of acid or acidic salt. This parameter change causes proteins to precipitate and separate, enabling bulk chromatographic isolation. The transformed egg white can then be processed through standard chromatography procedures, making large-scale production practical while maintaining purification quality.

Inventive Principle:
Principle #35Parameter changes

2Quantity of substance

If large volumes of egg white (at least 10 liters) are processed using traditional methods, then industrial-scale production is attempted, but costs and processing time increase significantly

Engineering Contradiction:
Improveegg white volumeVSAvoidprocessing time
Core Design Contradiction:
Quantity of substanceVSLoss of time

Solution Approach 1:

The patent applies preliminary acidification to egg white before chromatographic processing. By adjusting the pH to 2-4 in advance and allowing precipitation to occur, the protein separation is facilitated before the main chromatographic step. This preliminary action reduces the complexity and time of subsequent processing, making large-volume processing more efficient.

Inventive Principle:
Principle #10Preliminary action

3Ease of operation

If egg white is diluted to facilitate processing, then downstream isolation becomes easier, but the volume of material subject to processing increases, raising costs

Engineering Contradiction:
Improveprocessing easeVSAvoidmaterial volume
Core Design Contradiction:
Ease of operationVSQuantity of substance

Solution Approach 1:

Instead of diluting egg white, the patent changes its chemical parameter (pH) to induce protein precipitation and phase separation. This allows the process to proceed with concentrated egg white, reducing the volume of material requiring downstream processing while maintaining ease of operation through the chemical transformation.

Inventive Principle:
Principle #35Parameter changes

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 method significantly reduces the amount of egg white materials and processing time, minimizing costs and labor, while maintaining the integrity of therapeutic proteins, making large-scale production more efficient and cost-effective.

Implementation Method 1

adding an acidic buffer comprising an acidic agent to a pool of egg white... adjusting the pH of the egg white to from about 5 to about 6.5

Methodology Applied
Scientific EffectpH adjustment:

Implementation Method 2

allowing the mixed egg white to settle such that the egg white separates into top, middle and bottom layers

Methodology Applied
Scientific EffectPrecipitation: Precipitation

Implementation Method 3

filtering the egg white to form a solution (i.e., a clear solution)

Methodology Applied
Scientific EffectFiltration: Filter (physical)

Implementation Method 4

isolating the recombinant protein in the egg white by column chromatography

Methodology Applied
Scientific EffectChromatography: Chromatography

Data Source

PatentEP3133932B1Egg white processing
Publication Date: 2021.08.11 ALEXION PHARMACEUTICALS INC

AI summary

This disclosure relates to inexpensive and efficient methods of preparing egg white (e.g., obtained from eggs laid by transgenic chickens) for bulk chromatographic isolation of proteins (e.g., recombinant proteins) from the egg white, as well as methods of filtering acidified egg white and methods of isolating proteins from the egg white.