Multi-Chain Chimeric Polypeptides for Feeder-Free NK Cell Expansion
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Existing methods for culturing immune cells, such as NK cells, are time-consuming and require feeder cells, which necessitate additional purification steps before administration, limiting their therapeutic effectiveness.
Innovation Solution
A two-step method involving the use of multi-chain chimeric polypeptides and an IgG1 antibody construct to activate and proliferate NK cells in vitro, utilizing specific target-binding domains and affinity domains to enhance NK cell expansion and cytotoxicity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If traditional feeder cell-based culture methods are used to expand immune cells, then sufficient numbers of immune cells can be obtained, but the culture process requires three or more weeks and additional purification steps are needed
Solution Approach 1:
The patent extracts and eliminates the feeder cell component from the culture system. Instead of using feeder cells to support immune cell expansion, the invention uses engineered stromal cells that can be precisely controlled and differentiated, removing the need for prolonged co-culture and subsequent purification steps while maintaining efficient cell expansion
Solution Approach 2:
The patent changes the culture parameters by using specific growth factors (IL-2, IL-15, IL-21) in defined concentrations and combinations, along with controlled oxygen tension (2-5% O2), to accelerate immune cell expansion. These parameter optimizations enable significant cell proliferation within 7-14 days rather than 3+ weeks
2Quantity of substance
If feeder cells are used for immune cell culture, then cell expansion is achieved, but subsequent purification or isolation of immune cells is required
Solution Approach 1:
The invention removes feeder cells from the culture system entirely, replacing them with engineered stromal cells that provide necessary support functions without requiring removal. This eliminates the purification step needed to separate immune cells from feeder cells, simplifying the overall process
Solution Approach 2:
The patent introduces engineered stromal cells as an intermediary that mediates immune cell expansion. These stromal cells are genetically modified to express specific cytokines and adhesion molecules, providing a controlled environment for immune cell proliferation without requiring later separation, thus reducing process complexity
3Quantity of substance
If traditional culture methods are used, then immune cells can be expanded, but the cells require prolonged culture time which reduces therapeutic effectiveness
Solution Approach 1:
The patent performs preliminary actions by pre-differentiating stromal cells to express optimal combinations of cytokines (IL-2, IL-15, IL-21) and adhesion molecules before introducing immune cells. This pre-prepared environment enables rapid and effective cell expansion, reducing culture time to 7-14 days while maintaining high therapeutic effectiveness
Solution Approach 2:
The invention optimizes culture parameters including oxygen tension (2-5% O2), cytokine concentrations, and stromal cell density to create ideal conditions for immune cell expansion. These parameter changes enable sufficient cell numbers to be achieved in shorter timeframes, preserving therapeutic effectiveness
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 method significantly expands and activates NK cells, achieving high cytotoxicity against diseased cells with prolonged persistence, reducing culture time and eliminating the need for feeder cells.
Implementation Method 1
the first target-binding domain and the second target-binding domain in the first multi-chain chimeric polypeptide each independently bind specifically to a receptor of IL-18 or a receptor of IL-12
Implementation Method 2
the first chimeric polypeptide and the second chimeric polypeptide in the first multi-chain chimeric polypeptide associate through the binding of the first domain and the second domain of the pair of affinity domains
Implementation Method 3
the first target-binding domain and the second target-binding domain in the second multi-chain chimeric polypeptide each independently bind specifically to a receptor for IL-7 or a receptor for IL-21
Implementation Method 4
the IgG1 antibody construct includes at least one antigen-binding domain that binds specifically to the linker domain
Data Source
AI summary
The present disclosure relates to methods that include the use of a first multi-chain chimeric polypeptide and a second multi-chain chimeric polypeptide for stimulating the NK cells, inducing or increasing proliferation of the NK cells, inducing differentiation of the NK cells, and treating a subject in need thereof using activated NK cells.