Nonionic Surfactant Affinity Chromatography for HCP Removal
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Existing affinity chromatography methods for protein purification face challenges in effectively removing host cell proteins (HCPs) while maintaining high yield, as conventional additives for HCP removal can inadvertently reduce the recovery of target proteins.
Innovation Solution
Incorporating a nonionic surfactant, such as Tergitol 15-S series, into the protein sample before affinity chromatography to inhibit nonspecific binding of impurities, thereby enhancing the removal of host cell proteins without significantly affecting the recovery of target proteins.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If conventional additives are used to remove HCP during washing step, then HCP removal is improved, but antibody recovery is reduced due to nonspecific binding
Solution Approach 1:
A blocking agent is introduced as an intermediary substance that specifically binds to nonspecific binding sites on the Protein A resin and antibody molecules. This blocking agent prevents HCP from binding nonspecifically during the washing step, thereby improving HCP removal efficiency without causing loss of target antibody. The blocking agent acts as a mediator that selectively occupies unwanted binding sites while leaving specific Fc-Protein A binding sites intact.
Solution Approach 2:
The patent modifies the chemical environment by changing the composition of the washing buffer to include specific blocking agents (such as HSA, casein, or gelatin) at optimized concentrations. This parameter change in buffer composition alters the binding characteristics of the system, reducing nonspecific interactions between HCP and the chromatography matrix while maintaining specific antibody binding.
2Manufacturing precision
If washing step is performed to remove nonspecifically bound HCP, then impurity removal is improved, but process yield is reduced due to antibody loss
Solution Approach 1:
The blocking agent serves as an intermediary that preferentially occupies nonspecific binding sites on the Protein A resin and antibody molecules during the washing step. This allows the washing buffer to remove HCP without causing significant loss of specifically bound antibody, as the blocking agent prevents competing nonspecific interactions that would otherwise lead to antibody detachment.
Solution Approach 2:
The blocking agent is pre-added to the washing buffer before the washing step begins. This preliminary action ensures that blocking sites are occupied before HCP or antibody can engage in nonspecific binding, preventing unwanted interactions from occurring in the first place rather than attempting to reverse them afterward.
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 use of nonionic surfactants like Tergitol 15-S series effectively reduces nonspecific binding, improving the purity of target proteins by minimizing the loss of target proteins during the purification process while maintaining high yield.
Implementation Method 1
mixing a sample including a target protein with a composition including a nonionic surfactant
Data Source
Figure 1
Figure 2
Figure 3
AI summary
The present disclosure relates to a composition for improving the removal of impurities in protein purification by affinity chromatography, the composition including a nonionic surfactant, and to a method of purifying a target protein by using the composition. The composition or method according to an aspect, when used to purify a target protein, can effectively remove HCP while improving the yield of the target protein and thus may be applicable to manufacturing processes for recombinant protein pharmaceuticals such as antibody therapeutics.