Apatite Surface Neutralization for Chromatography Reuse
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Solution Overview
Problem
Current methods for purifying biomolecules using apatite solid supports often result in the release of hydronium ions, which can harm the target molecule and degrade the apatite surface, limiting the reuse of the material.
Innovation Solution
A method involving the use of a neutralization solution with amino compounds or sulphonated amines, in combination with alkali metal or alkali earth ions, to neutralize the apatite surface without eluting the target molecule, followed by a different elution solution to purify the molecule.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If a neutralization solution is used to neutralize the apatite surface, then the integrity of the apatite surface is maintained and reuse is enabled, but there is a risk of eluting the target molecule
Solution Approach 1:
The patent applies parameter changes by carefully controlling the pH range (6.5-9.0) and ionic strength (100 mM or less of alkali metal/alkali earth ion) of the neutralization solution. These parameter adjustments enable surface neutralization while preventing target molecule elution, resolving the contradiction between maintaining apatite integrity and retaining the target molecule.
Solution Approach 2:
The neutralization solution acts as an intermediary between the apatite surface and the target molecule. It contains basic amino compounds or sulphonated amine compounds that neutralize surface charge without directly interacting with or eluting the target molecule, thus mediating the neutralization process while preserving target retention.
2Productivity
If conventional elution methods are used, then the target molecule can be recovered, but the apatite surface degrades and cannot be reused
Solution Approach 1:
The patent applies preliminary action by performing surface neutralization before the elution step. The apatite surface is pre-treated with a neutralization solution to adjust its charge state, which enables subsequent efficient elution of the target molecule while preventing surface degradation, thus extending the support lifetime.
Solution Approach 2:
The patent segments the purification process into distinct steps: (1) adsorption of target molecule to apatite, (2) neutralization of apatite surface with controlled solution, and (3) elution with a different solution. This segmentation allows optimization of each step independently, enabling both target recovery and surface preservation.
3Reliability
If the neutralization solution has high ionic strength, then the apatite surface can be effectively neutralized, but the target molecule elutes from the surface
Solution Approach 1:
The patent applies parameter changes by optimizing the ionic strength to 100 mM or less of alkali metal or alkali earth ion in the neutralization solution. This parameter control enables sufficient surface neutralization while maintaining target molecule adsorption, resolving the contradiction between neutralization efficiency and target retention.
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 prevents significant elution of the target molecule during neutralization, maintains the integrity of the apatite surface, and allows for multiple cycles of purification without degrading the material.
Implementation Method 1
contacting the solid surface comprising the adsorbed target molecule with a solution comprising a basic amino compound and alkali metal ion or alkali earth ion, or a sulphonated amine compound and alkali metal ion or alkali earth ion, sufficient to neutralize the apatite solid surface
Implementation Method 2
contacting a sample comprising the target molecule to an apatite solid surface thereby adsorbing the target molecule to the solid surface
Implementation Method 3
eluting the target molecule from the solid support by contacting the solid support with a solution different from the solution in step (b)
Data Source
AI summary
The present invention discloses methods of neutralizing apatite surfaces, for example during chromatography and before protein elution.