Protein Viscosity Reduction via HDX-MS Region Identification
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Solution Overview
Problem
Highly concentrated therapeutic monoclonal antibodies exhibit high viscosity, leading to increased injection time, pain, and manufacturing costs, as well as challenges in bioprocessing, due to short-range electrostatic and hydrophobic protein-protein interactions, which existing methods struggle to characterize and address effectively.
Innovation Solution
The method involves microdialysis of protein samples against a deuterium-containing buffer followed by hydrogen/deuterium exchange mass spectrometry to identify regions contributing to viscosity, allowing for modification of these regions to reduce or increase viscosity as needed, using techniques such as amino acid substitution.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If therapeutic antibodies are formulated at high concentration for subcutaneous injection, then the dosing frequency is reduced and patient convenience is improved, but the viscosity of the formulation increases causing increased injection time and pain
Solution Approach 1:
The patent modifies the amino acid sequence of the therapeutic antibody by introducing specific mutations (e.g., in the CDR regions) to change the biophysical parameters of the protein. These sequence changes alter the protein-protein interaction characteristics, enabling the antibody to maintain low viscosity even at high concentrations (e.g., 50-200 mg/mL), thus resolving the contradiction between high dosing concentration and low injection time
Solution Approach 2:
The patent focuses modifications on specific local regions of the antibody molecule, particularly the complementarity determining regions (CDR1, CDR2, CDR3) of the heavy and light chains. By making localized changes only in these specific regions rather than the entire protein, the invention achieves viscosity reduction while preserving the overall therapeutic activity and specificity of the antibody
2Productivity
If therapeutic antibodies are formulated at high concentration, then manufacturing efficiency is improved, but the viscosity increases causing destabilization and increased manufacturing costs
Solution Approach 1:
The patent changes the amino acid sequence parameters of the antibody by introducing specific mutations that alter the electrostatic and hydrophobic interaction characteristics. These parameter changes prevent harmful protein-protein interactions that would otherwise occur at high concentrations, thereby maintaining formulation stability while enabling concentrated manufacturing and storage
3Measurement precision
If conventional structural techniques are used to characterize antibody conformation, then detailed structural analysis can be obtained, but the methods require very specialized skills, large amounts of sample, and have long turnaround time
Solution Approach 1:
The patent replaces complex mechanical and computational structural analysis methods with a simpler biochemistry-based approach using hydrogen/deuterium exchange mass spectrometry (HDX-MS). This substitution maintains the ability to characterize protein structure and dynamics while dramatically reducing the required expertise, sample amount, and analysis time
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 enables the identification and modification of viscosity-contributing regions in proteins, thereby reducing high viscosity issues in concentrated protein formulations, improving injection processes and manufacturing efficiency.
Implementation Method 1
microdialysing samples of the protein in a microdialysis cartridge against a buffer containing deuterium
Implementation Method 2
hydrogen/deuterium exchange mass spectrometry to determine regions of the protein in the sample that have reduced levels of deuterium
Data Source
AI summary
Systems and methods for determining regions of proteins that contribute to self-association of the protein are provided. Methods for modifying the self-association of concentrated protein formulations are also provided.