Squalene Purification from Yeast Using Extraction and Chromatography
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Solution Overview
Problem
Existing methods struggle to produce squalene from renewable resources with high purity and in a sustainable manner, particularly due to the challenge of purifying squalene from intracellularly sequestered host cells like yeast, where it is not secreted in appreciable amounts.
Innovation Solution
A method involving extraction, evaporation, and chromatography steps to purify squalene from squalene-producing host cells, utilizing homogenization, centrifugation, solvent extraction, demulsification, and chromatography techniques such as alumina or silica chromatography to achieve high purity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If squalene is produced from host cells through fermentation, then renewable and sustainable production is achieved, but purification from intracellular impurities becomes difficult
Solution Approach 1:
The purification process is divided into distinct sequential steps: cell lysis, extraction, evaporation, and chromatography. Each step targets specific impurities at different stages, making the overall purification of intracellular squalene feasible and effective.
Solution Approach 2:
The patent employs extraction techniques to separate squalene from intracellular components. By using appropriate solvents and extraction methods, squalene is selectively removed from the complex intracellular matrix, enabling high-purity production from renewable host cells.
2Ease of operation
If host cells are homogenized to release intracellular squalene, then squalene accessibility is improved, but complexity of impurity matrix increases
Solution Approach 1:
After homogenization releases squalene into the extracellular media along with impurities, extraction techniques are applied to selectively remove squalene from the complex matrix. This separates the target compound from the released impurities, managing the complexity introduced by homogenization.
Solution Approach 2:
The patent uses intermediary substances such as solvents and chromatography media to facilitate separation. These intermediaries interact selectively with squalene and impurities, enabling clean separation from the complex homogenized mixture without requiring direct complex processing of the entire matrix.
3Manufacturing precision
If multiple purification steps are implemented, then squalene purity is improved, but process complexity and time increase
Solution Approach 1:
The purification process is segmented into essential steps (extraction, evaporation, chromatography), where each step performs a specific function. This segmentation allows for optimized timing and efficiency, achieving high purity without unnecessary delays in any single step.
Solution Approach 2:
The patent ensures continuous and efficient progression through purification steps, minimizing idle time between operations. By maintaining continuous useful action throughout the process, high purity is achieved while optimizing the overall time required for purification.
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
The method achieves squalene compositions with unprecedented purity levels, addressing the challenges of impurity removal and enhancing the sustainability of squalene production from renewable sources.
Implementation Method 1
The extraction step includes one or more of: homogenizing the fermentation composition, (ii) solvent extraction of the fermentation composition
Implementation Method 2
The method may further include evaporating the squalene resulting from the extraction step
Implementation Method 3
purifying the squalene resulting from the evaporation step by way of chromatography, such as an alumina or a silica chromatography process
Implementation Method 4
separating the homogenized fermentation composition resulting from (i) into sediment and supernatant by way of centrifugation
Data Source
AI summary
The present disclosure provides compositions and methods for producing high-purity squalene from a squalene source, for example from host cells, such as yeast cells that are capable of synthesizing squalene. Further provided herein are compositions containing high-purity squalene, as well as methods of using the same in pharmaceutical formulations, such as in adjuvant formulations for use in vaccines.