Antibody-Producing Cell Immortalization While Preserving Secretion
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Solution Overview
Problem
Existing methods fail to effectively immortalize antibody-producing cells while retaining their antibody-producing ability, particularly when using genes like Bcl-2, Myc, and Cond1, or c-Myc and SV40 large T antigen, as they either do not confer immortalization or impair the desired traits.
Innovation Solution
Introduce SV40 T antigen gene, Bcl-2 family genes, and one or more genes from the Myc and cyclin D family into antibody-producing cells to achieve transformation and immortalization, maintaining antibody-producing capability.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Duration of action of stationary object
If Bcl-2 gene, Myc gene, and Cond1 gene are introduced into antibody-producing cell to perform transformation, then the cell can acquire immortalization ability, but the cell cannot retain the antibody-producing ability
Solution Approach 1:
The invention changes the genetic parameters by introducing a specific combination of four genes (SV40 T antigen, Bcl-2, Myc, and Cond1) rather than using traditional transformation methods. This parameter change enables the antibody-producing cell to acquire immortalization ability while surprisingly retaining its antibody-producing capability, resolving the contradiction between extended cell division capacity and functional reliability
Solution Approach 2:
The invention uses a composite genetic approach by combining multiple cancer-related genes (SV40 T antigen, Bcl-2, Myc, and Cond1) in a specific configuration. This composite genetic material works synergistically to achieve both immortalization and preservation of antibody-producing function, which cannot be achieved by introducing individual genes or traditional gene combinations alone
2Speed
If c-Myc gene and SV40 large T antigen gene are introduced into antibody-producing cell to perform transformation, then the cell can acquire proliferative ability, but the useful traits cannot be retained
Solution Approach 1:
The invention modifies the transformation parameters by including Bcl-2 and Cond1 genes in addition to SV40 T antigen and Myc genes. This parameter adjustment creates a balanced genetic configuration that promotes cell proliferation while simultaneously preserving the antibody-producing phenotype, overcoming the limitation of traditional two-gene transformation approaches
3Reliability
If hybridoma method is used for production of monoclonal antibodies in rodents, then immortalized cells with antibody-producing ability can be obtained, but the method is not applicable to non-rodent species requiring complicated genetic engineering
Solution Approach 1:
The invention creates a universal transformation system using four cancer-related genes that can be applied across different species including human, rodent, and non-rodent species. This multi-functional genetic approach replaces species-specific hybridoma methods, enabling immortalization of antibody-producing cells from any species through a standardized protocol without requiring complicated species-adapted genetic engineering
Solution Approach 2:
The invention segments the immortalization process into four distinct genetic components (SV40 T antigen, Bcl-2, Myc, and Cond1 genes), each contributing a specific function. This segmentation allows for modular application and optimization across different species while maintaining the overall effectiveness of the transformation process, simplifying the genetic engineering requirements compared to traditional hybridoma methods
Data Source
AI summary
An object of the present invention is to provide a method for immortalizing an antibody-producing cell; and an immortalized transformed cell obtained by the method. The invention relates to a method for producing an immortalized cell, the method including introducing SV40 T antigen gene, genes of Bcl-2 family, and one or more genes selected from the group consisting of genes of Myc family and genes of cyclin D family into a non-immortalized antibody-producing cell to produce an immortalized cell; and a method for producing an immortalized antibody-producing cell, the method including collecting an antibody-producing cell from a mammal and subjecting the cell to primary culture; and introducing one or more genes selected from the group consisting of SV40 T antigen gene, genes of Bcl-2 family, genes of Myc family, and genes of cyclin D family into a primary antibody-producing cell thus obtained to produce an immortalized antibody-producing cell.