Immortalized Human B Cells for Monoclonal Antibody Production
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Solution Overview
Problem
Current methods for producing human monoclonal antibodies are inefficient and often result in antibodies that are not fully human, leading to immune responses and reduced therapeutic efficacy due to the use of murine sequences, and the process of obtaining stable human antibody-secreting cell lines is complex and time-consuming.
Innovation Solution
A method involving the selection, stimulation, and immortalization of antibody-secreting cells based on cell surface markers and specific agents, followed by exposure to a viral immortalizing agent, to generate polyclonal populations of cells that secrete specific isotypes, allowing for high-throughput analysis and storage for future screening.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Duration of action of stationary object
If hybridoma technology is used to produce human monoclonal antibodies, then continuous production is achieved, but the process becomes complex and time-consuming due to fusion requirements and instability issues
Solution Approach 1:
The patent extracts the essential function of continuous antibody production from the complex hybridoma fusion process. By using immortalized B cells that can be directly transduced with antigen-specific receptors, the method eliminates the need for cell fusion and hybridoma formation, achieving continuous production through a simpler transduction-based approach
Solution Approach 2:
The patent segments the antibody production process into distinct modular steps: immortalization of B cells, transduction with antigen-specific receptors, and selection. This modular approach replaces the integrated but complex hybridoma fusion process, allowing each step to be optimized independently while maintaining continuous production capability
2Productivity
If murine antibody sequences are used, then production efficiency is improved, but immune responses are triggered reducing therapeutic efficacy
Solution Approach 1:
The patent changes the fundamental parameter of antibody origin from murine to fully human by using human B cells as the starting material. The immortalized human B cells are transduced with human antigen-specific receptors, ensuring that the produced antibodies are fully human in sequence and structure, thereby eliminating immune response while maintaining production efficiency
Solution Approach 2:
The patent introduces immortalized human B cells as an intermediary system that can be transduced with antigen-specific receptors. This intermediary approach allows the generation of fully human antibodies through transduction rather than direct immunization, achieving both high productivity and full human compatibility
3Reliability
If primary antibody-secreting cells are used, then full human specificity is achieved, but cell viability and proliferation are insufficient for high-throughput screening
Solution Approach 1:
The patent applies preliminary immortalization treatment to human B cells before transduction with antigen-specific receptors. This preliminary action of immortalization ensures that the cells will proliferate continuously in culture, providing sufficient cell numbers for high-throughput screening while maintaining full human antibody specificity
Solution Approach 2:
The patent establishes continuous proliferation of human B cells through immortalization, allowing the useful action of antibody secretion to continue indefinitely. This continuous cell population maintains full human specificity while providing the productivity needed for high-throughput screening and antibody production
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 enhances cell viability and proliferation, resulting in a larger diversity and number of immortalized antibody-secreting cells that can be screened directly, expediting the identification and production of monoclonal antibodies with desired antigen specificity and biological activity.
Implementation Method 1
exposing said population of selected and stimulated cells to the immortalizing agent in cell culture conditions
Data Source
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AI summary
The present Invention provides novel methods for immortalizing cells that secrete antibodies of one or more specific isotypes. Polyclonal, oligoclonal, and monoclonal populations of cells obtained using the methods of the Invention can be screened on the basis of the functional and/or binding activities of the antibodies they secrete, for example directed to antigens of human or viral origin having medical interest, in cell culture conditions. Using these methods, human B cells that secrete antibodies binding human Cytomegalovirus, Herpes Simplex Virus, or HSP60 protein have been efficiently immortalized with Epstein-Barr virus.