Expression Vector with MAR and LCR for High Protein Yield
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for producing therapeutic recombinant proteins and antibodies in animal cells face inefficiencies in gene expression and productivity, necessitating the development of more effective expression vectors to enhance protein production levels and stability.
Innovation Solution
An expression vector is designed incorporating a matrix attachment region (MAR) element, a simian virus 40 (SV40) promoter operably linked to a chimeric intron, and locus control regions (LCRs), specifically optimized for use in CHO cells to significantly increase gene expression levels and protein production.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional expression vectors are used for protein production in animal cells, then the production process can be established, but the gene expression level and productivity remain low
Solution Approach 1:
The expression vector is segmented into distinct functional modules: MAR element for chromatin organization and stable transcription, SV40 promoter for strong gene expression initiation, chimeric intron for enhanced mRNA processing and stability, and LCR for high-level expression control. Each module performs a specific function that collectively resolves the contradiction between productivity and reliability.
Solution Approach 2:
The vector constructs a composite genetic system by combining heterogeneous elements from different sources: MAR elements from chromatin studies, SV40 promoter from viral systems, chimeric introns with splice sites, and LCR from high-expression loci. This composite structure integrates multiple mechanisms that simultaneously enhance both expression levels and stability.
2Productivity
If gene amplification is performed to increase expression levels, then productivity improves, but the time required for cell line development increases
Solution Approach 1:
The vector incorporates all necessary elements for high-level expression (MAR, promoter, intron, LCR) in the initial construction, eliminating the need for subsequent gene amplification steps. This preliminary configuration of optimal expression elements allows direct selection of high-producing cell lines, reducing development time from months to weeks while maintaining high productivity.
3Productivity
If multiple vector optimization elements are added to enhance expression, then productivity increases, but vector complexity increases
Solution Approach 1:
Each element in the vector serves multiple functions: the MAR element provides both chromatin organization and stable transcription; the chimeric intron contributes to both splicing efficiency and mRNA stability; the LCR enhances both expression level and consistency. This multi-functionality reduces the need for additional separate elements, managing complexity while maximizing productivity.
Data Source
Figure 1
Figure 2~3
Figure 4
AI summary
The present invention relates to an expression vector having an improved ability to express a gene, cells transformed by the expression vector, and a method for mass-producing a target protein by using the cells. The expression vector according to the present invention shows a more excellent ability to express a gene than a typical animal cell expression vector, and thus, protein expression of a heterogeneous gene can be significantly increased.