Recombinant Transition Vector for High-Level Protein Expression
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Solution Overview
Problem
Current baculovirus expression vector systems using insect cells for foreign protein production face low expression levels due to inefficient signal sequence recognition and protein processing, particularly when using only the polyhedrin gene promoter, leading to the need for costly and inefficient purification processes.
Innovation Solution
A recombinant transition vector combining a polyhedrin protein coding gene promoter with a homologous region 5 (hr5) sequence and a vp39 promoter, along with a burst sequence, to enhance foreign protein expression in insect cells, allowing for high-level production of target proteins in their native form without fusion with other proteins.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Device complexity
If only the polyhedrin gene promoter is used for foreign protein expression in insect cells, then the expression system is simple, but the protein expression level is low
Solution Approach 1:
The patent combines multiple promoters (polyhedrin promoter, vp39 promoter, and hr5 sequence) into a single recombinant transition vector to achieve synergistic enhancement of foreign protein expression. This merging of functional elements resolves the contradiction by increasing productivity while maintaining manageable complexity through integrated design.
Solution Approach 2:
The recombinant transition vector functions as a composite genetic system integrating multiple promoter elements (polyhedrin promoter, vp39 promoter, hr5 sequence) that work together to enhance transcriptional activity. This composite approach enables high protein expression levels while maintaining a unified vector structure.
2Quantity of substance
If the foreign protein is fused with partial polyhedrin protein to increase expression amount, then the protein production quantity increases, but additional cleavage and purification processes are required
Solution Approach 1:
The patent extracts and eliminates the need for fusion with polyhedrin protein by using the hr5 sequence and vp39 promoter to drive direct high-level expression of the foreign protein. This extraction of the fusion requirement resolves the contradiction by achieving high protein quantity without the necessary cleavage and purification steps.
Solution Approach 2:
The patent converts the potential harm of requiring fusion (which necessitates cleavage and purification) into a benefit by using the hr5 sequence and vp39 promoter to achieve high expression without fusion. This approach transforms the processing complexity into a simplified direct expression system.
3Productivity
If the foreign protein is expressed using a strong promoter of polyhedrin protein coding gene, then the expression system is efficient, but the expression level of foreign protein is smaller compared with original polyhedrin protein
Solution Approach 1:
The patent changes the promoter parameters by introducing the hr5 sequence and vp39 promoter in addition to the polyhedrin promoter. This parameter enhancement resolves the contradiction by achieving both high expression efficiency and high foreign protein expression levels through synergistic promoter action.
Solution Approach 2:
The recombinant transition vector creates a composite promoter system combining polyhedrin promoter, vp39 promoter, and hr5 sequence that works together to overcome the limitation of single promoter systems. This composite approach enables the foreign protein to be expressed at levels comparable to or exceeding the original polyhedrin protein while maintaining expression efficiency.
Data Source
AI summary
Disclosed are a novel recombinant transition vector for increasing expression of a foreign protein in a native form without fusion partners, and a method for mass production of a foreign target protein using the same. The recombinant transition vector according to the present disclosure may allow a large amount of a foreign target protein with a high therapeutic and prophylactic value to be expressed in an insect cell. In particular, the vector may increase the expression of the foreign target protein in an own form thereof, not fused with other fusion partners. Therefore, the use of the recombinant transition vector may produce useful proteins such as antigens in insect cells at low cost and high efficiency.