Antibody Formulation Stability Against Aggregation
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Solution Overview
Problem
There is a need for a shear and temperature stable pharmaceutical formulation of antibodies, particularly anti-CD20 antibodies, that can maintain biological activity and structural integrity during storage and therapeutic use, as existing formulations face challenges with chemical and physical instability such as aggregation, deamidation, and oxidation.
Innovation Solution
A stable aqueous antibody formulation comprising ofatumumab with specific concentrations of sodium acetate, sodium chloride, arginine free base, EDTA, and polysorbate 80, adjusted to a pH range of 5.0 to 7.0, which provides stability at various temperatures and under stress conditions like shaking and elevated temperatures.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional antibody formulations are used, then the formulation can be stored and administered, but the antibody suffers from chemical and physical instability including aggregation, deamidation, and oxidation
Solution Approach 1:
The formulation uses specific concentration ranges of excipients (sodium acetate 10-100 mM, sodium chloride 25-100 mM, arginine 0.5-5%, EDTA 0.02-0.2 mM, polysorbate 80 0.01-0.2%) and pH adjustment (5.0-7.0) to optimize antibody stability. These parameter optimizations prevent degradation pathways while maintaining solubility and biological activity
Solution Approach 2:
Multiple excipients act as intermediaries to protect the antibody: sodium acetate and sodium chloride maintain ionic strength and pH buffer capacity, arginine prevents deamidation and aggregation, EDTA chelates metal ions to prevent oxidation, and polysorbate 80 prevents surface adsorption and aggregation. Each excipient mediates specific protective functions
2Adaptability or versatility
If the formulation is exposed to stress conditions like elevated temperature and shaking, then therapeutic flexibility is improved, but the antibody undergoes denaturation and aggregation
Solution Approach 1:
The formulation includes excipients that provide preemptive protection against stress conditions: arginine cushiones against thermal denaturation and shear-induced aggregation, polysorbate 80 cushions against surface adsorption during shaking, and the buffer system cushions against pH shifts during storage. This prior cushioning enables the formulation to withstand stress conditions like elevated temperature and shaking without significant degradation
3Quantity of substance
If the formulation maintains high concentration for effective therapy, then therapeutic efficacy is improved, but the antibody is more prone to aggregation and precipitation
Solution Approach 1:
The formulation optimizes excipient concentrations specifically to enable high antibody concentrations without aggregation: arginine at 0.5-5% maintains solubility and prevents aggregation even at high antibody concentrations, while the combination of sodium acetate, sodium chloride, and polysorbate 80 creates an environment that suppresses aggregation pathways. This parameter optimization allows therapeutic concentrations to be achieved while maintaining stability
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 formulation achieves long-term stability, retaining biological activity and preventing aggregation and denaturation, with the ability to maintain clarity and prevent precipitation even under stress conditions, thus ensuring effective therapeutic use for extended periods.
Implementation Method 1
Chemical instability can result from deamidation, racemization, hydrolysis, oxidation, beta elimination or disulfide exchange
Implementation Method 2
Chemical instability can result from deamidation, racemization, hydrolysis, oxidation, beta elimination or disulfide exchange
Implementation Method 3
Physical instability can result from denaturation, aggregation, precipitation or adsorption, for example
Implementation Method 4
The formulation further comprises 10 to 100 mM sodium acetate, 25 to 100 mM sodium chloride, 0.5 to 5% arginine free base, 0.02 to 0.2 mM EDTA, 0.01 to 0.2% polysorbate 80 and adjusted to pH 5.0 to 7.0
Implementation Method 5
Proteins are larger and more complex than traditional organic and inorganic drugs (i.e. possessing multiple functional groups in addition to complex three-dimensional structures)
Implementation Method 6
For a protein to remain biologically active, a formulation must preserve the intact conformational integrity of at least a core sequence of the protein's amino acids
Data Source
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Figure 3
AI summary
This invention relates to a shear and temperature stable antibody formulations that are more stable than compared to a standard formulation (such as 30 mM citrate, 100mM NaCl, pH 6.5). The present invention's shear and temperature stable antibody formulations show reduced precipitation when subjected to stress conditions but the standard formulation had aggregated. This result was unpredictable because thermodynamically the two formulations are similar as seen by their DSC (differential scanning calorimeter) profiles.