Coiled Coil and Tether Protein Complexes for Multispecific Antibody Assembly
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Solution Overview
Problem
Current methods for producing multispecific antibodies face challenges such as poor solubility, low yield, immunogenicity, and instability, making them unsuitable for commercial and therapeutic applications.
Innovation Solution
The development of novel protein complexes involving coiled coil domains linked to Fc CH components, with optional cleavability, and the use of tethers to form stable protein complexes, allowing for efficient production of homogenous heteromultimeric complexes through specific nucleic acid sequences and enzymatic reactions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional methods are used to produce multisspecific antibodies, then production processes exist, but the products suffer from poor solubility, low yield, immunogenicity, and instability
Solution Approach 1:
The invention divides the antibody molecule into separate functional components: a first polypeptide with a coiled coil domain for heterodimerization, a second polypeptide with an Fc CH component, and optional tethers. This segmentation allows each component to be optimized independently for stability and yield, while assembling into a functional multispecific antibody complex.
Solution Approach 2:
The coiled coil domain acts as an intermediary element that mediates the association between different polypeptide chains. It provides a controlled interface for heterodimerization, ensuring stable and specific binding between the first and second polypeptides while preventing homodimerization, thereby improving both stability and yield.
2Productivity
If conventional antibody production methods are used, then multispecific antibodies can be produced, but they demonstrate low yield of heterodimer formation and are technically challenging to manufacture
Solution Approach 1:
The invention introduces local structural features at specific locations: a coiled coil domain with specific heptad repeat patterns at the N-terminus of the first polypeptide, and a Fc CH component at the C-terminus. These localized structural modifications create specific binding interfaces that promote heterodimer formation while simplifying the overall manufacturing process.
Solution Approach 2:
The invention modifies key parameters of the protein structure, including the introduction of cleavable tethers with specific enzymatic cleavage sites and the optimization of coiled coil domain sequences. These parameter changes enable controlled assembly and purification, reducing manufacturing complexity while increasing heterodimer yield.
3Manufacturing precision
If tethers are used to link polypeptides, then proper association of light and heavy chains is achieved, but the tethers may require enzymatic cleavage and add complexity
Solution Approach 1:
The tether system is designed to be dynamic rather than static. The tethers are cleavable through enzymatic reactions, allowing them to transition from a linked state during production to a separated state for purification. This dynamic design achieves precise chain association during manufacturing while enabling simplified processing afterward.
Solution Approach 2:
The tethers are designed as temporary, disposable elements that fulfill their purpose during protein assembly and then are discarded through enzymatic cleavage. This approach achieves the necessary manufacturing precision for proper chain association without permanently adding complexity to the final product structure.
4Reliability
If coiled coil domains are used to prevent homodimerization, then heterodimer formation is promoted, but the domains must be engineered with specific sequences
Solution Approach 1:
The coiled coil domains are designed with asymmetric sequences that prevent homodimerization. The first polypeptide contains a specific coiled coil sequence that is different from any potential homologue, creating an asymmetric binding interface that only accommodates the complementary second polypeptide. This asymmetry ensures reliable heterodimer formation while preventing homodimerization.
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 high-yield, stable production of multispecific protein complexes, such as antibodies, that can bind multiple targets, facilitating therapeutic applications by ensuring proper association of light and heavy chains and reducing homodimerization of heavy chains.
Implementation Method 1
coiled coil domains that are linked to an Fc CH component
Implementation Method 2
stable protein complexes
Implementation Method 3
tether and an Fc CH component complex, which tether may or may not be cleavable from the protein
Implementation Method 4
the tether is cleaved by a host cell or cleaved by a chemical or enzymatic reaction in vitro
Data Source
AI summary
The invention provides engineered protein complexes constructed using a coiled coil and/or a tether and methods for making, using, and purifying such complexes, such as multispecific antibodies or other multispecific Fc containing complexes.