HIC Column Regeneration Using Low-Conductivity Alkaline Cleaning
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Solution Overview
Problem
Existing hydrophobic interaction chromatography (HIC) media regeneration methods are inefficient in removing residual materials, leading to contamination and reduced longevity, and often require lengthy cleaning processes that may introduce additional concerns.
Innovation Solution
A method involving the use of an alkaline solution with a pH between 10 and 14 and conductivity of 0.5 mS/cm to 10 mS/cm is applied to HIC media to remove bound materials, utilizing sodium hydroxide, potassium hydroxide, calcium hydroxide, or Tris, with a total dissolved salt concentration between 0.1 mM and 10 mM, and optionally followed by a chaotropic agent to achieve complete regeneration.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional HIC media regeneration methods are used, then the media can be reused for multiple cycles, but residual materials are not effectively removed leading to contamination and reduced longevity
Solution Approach 1:
The patent changes the chemical parameters of the regeneration solution by specifying pH between 10-14 and conductivity between 0.5-10 mS/cm, which creates optimal conditions for removing residual materials while preserving media integrity and enabling effective media regeneration across multiple cycles
Solution Approach 2:
The patent introduces an alkaline solution as an intermediary substance with specific pH and conductivity characteristics that mediates between the bound materials and the HIC media, effectively removing contaminants without directly damaging the media structure
2Object-generated harmful factors
If lengthy cleaning processes are used to remove residual materials, then contamination is reduced, but the process time increases and additional concerns are introduced
Solution Approach 1:
The patent applies preliminary action by using an alkaline solution that pre-conditioned the media for effective regeneration, removing residual materials in a single step before subsequent chaotropic agent treatment, thereby reducing overall process time while maintaining thorough cleaning
Solution Approach 2:
The patent skips intermediate cleaning steps by directly applying the alkaline solution with optimized pH and conductivity parameters that achieve effective residual material removal in one pass, eliminating the need for multiple sequential cleaning operations
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 effectively reduces residual mass on HIC media to less than 1.0%, preserving media quality and extending its lifespan without the need for extensive cleaning, while maintaining the integrity of the chromatography process.
Implementation Method 1
hydrophobic interaction chromatography (HIC) may separate molecules according to differences in surface hydrophobicity, by using a reversible interaction between the molecules and hydrophobic surfaces of a HIC medium
Implementation Method 2
passing one or more column volumes of an alkaline solution through hydrophobic interaction media within the column, wherein the alkaline solution exhibits a pH of between 10 and 14, and a conductivity of between 0.5 mS/cm and 10 mS/cm, wherein material bound to the hydrophobic interaction media is removed
Data Source
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AI summary
Aspects of the present disclosure relate to a method of regenerating a hydrophobic interaction chromatography column to which a load mass has been applied, the method comprising passing one or more column volumes of an alkaline solution through hydrophobic interaction media within the column, wherein the alkaline solution exhibits a pH of between about 10 and about 14, and a conductivity of between 0.5 mS/cm and about 10 mS/cm, wherein material bound to the hydrophobic interaction media is removed. In some cases, the alkaline solution may include sodium hydroxide at a concentration of between, e.g., about 0.1 mM and 10 mM.