Four-Gene Immortalization Preserving Antibody-Producing Cell Function
Find Innovative SolutionsGenerate Solutions
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 Ccnd1, and introducing c-Myc and SV40 large T antigen genes results in loss of useful traits.
Innovation Solution
Introduce SV40 T antigen gene, Bcl-2 family genes, and one or more genes from the Myc and cyclin D families into antibody-producing cells to achieve immortalization while maintaining antibody production capability.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Duration of action of stationary object
If Bcl-2 gene, Myc gene, and Ccnd1 gene are introduced into antibody-producing cells to perform transformation, then the cells acquire immortalization ability, but the cells 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 Ccnd1) rather than the conventional three genes. This parameter change in the genetic composition enables the cells to simultaneously achieve immortalization and retain antibody-producing ability, resolving the contradiction between extended cell lifespan and functional reliability
Solution Approach 2:
The invention creates a composite genetic structure by combining four specific genes working together in the antibody-producing cells. This composite approach, where multiple genetic elements are introduced as a integrated set, produces a synergistic effect that allows both immortalization and preservation of antibody-producing function, which cannot be achieved by individual genes or smaller combinations
2Productivity
If c-Myc gene and SV40 large T antigen gene are introduced into antibody-producing cells to perform transformation, then the cells acquire proliferative ability, but the useful traits are lost
Solution Approach 1:
The invention modifies the genetic parameters by adding Bcl-2 and Ccnd1 genes to the conventional c-Myc and SV40 large T antigen combination. This parameter expansion creates a more balanced genetic profile that promotes proliferation through Myc and Ccnd1 while Bcl-2 prevents apoptosis, thereby maintaining cell viability and useful traits during rapid division
Solution Approach 2:
The SV40 T antigen acts as an intermediary that facilitates the interaction between the introduced genes and the cell's endogenous regulatory mechanisms. It mediates the transformation process by binding to retinoblastoma protein and releasing E2F transcription factors, thereby enabling cell cycle progression while allowing the cell to maintain its differentiated functions including antibody production
Data Source
Figure 1~2
Figure 3~4
Figure 5~6
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.