Ebola Vaccine Codon Optimization Adenovirus Vector

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Solution Overview

Problem

Current Ebola vaccines lack high efficiency in preventing the disease, and the AdMax system requires optimization to enhance the expression of the Ebola virus glycoprotein antigen for improved immune response without increasing immune dosage or adverse effects.

Innovation Solution

Codon optimization of the Ebola virus glycoprotein gene to increase its expression level in eukaryotic cells, using a replication-defective human adenovirus vector packaged with the AdMax system, which results in a recombinant adenovirus vaccine capable of inducing robust humoral and cellular immune responses.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If conventional inactivated vaccines or sub-unit vaccines are used, then the vaccine structure is simple and easy to manufacture, but they are ineffective against Ebola virus

Engineering Contradiction:
Improvevaccine effectivenessVSAvoidmanufacturing complexity
Core Design Contradiction:
ReliabilityVSEase of manufacture

Solution Approach 1:

The patent uses a replication-defective adenovirus vector as an intermediary carrier to deliver the Ebola virus GP antigen gene into host cells. This vector system enables effective antigen expression and immune response induction without requiring complex inactivated virus preparation or sub-unit purification processes, thus resolving the contradiction between effectiveness and ease of manufacture.

Inventive Principle:
Principle #24Intermediary (Mediator)

Solution Approach 2:

The patent employs codon optimization to change the nucleotide sequence parameters of the GP antigen gene, optimizing it for mammalian cell expression. This parameter change enables high-level antigen production from the viral vector, significantly improving vaccine effectiveness while maintaining the relative simplicity of viral vector manufacturing.

Inventive Principle:
Principle #35Parameter changes

2Reliability

If more GP antigen is provided to enhance immune response, then the protective effect is improved, but the immune dosage increases and adverse effects occur

Engineering Contradiction:
Improveprotective effectVSAvoidadverse effects
Core Design Contradiction:
ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The patent optimizes the GP antigen gene through codon usage optimization tailored for mammalian cells, which dramatically enhances the translation efficiency and antigen expression levels. This allows sufficient antigen production from a low viral dosage, achieving strong protective effect without increasing the immune dosage and causing adverse effects.

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The replication-defective adenovirus vector enters host cells and uses the host's own cellular machinery to produce the GP antigen. This self-service mechanism within host cells ensures efficient antigen production with minimal external dosage, avoiding the adverse effects associated with high antigen dosing.

Inventive Principle:
Principle #25Self-service

3Productivity

If the AdMax system is used for adenovirus packaging, then the virus production efficiency is improved, but the expression level of Ebola GP antigen needs further optimization

Engineering Contradiction:
Improvevirus production efficiencyVSAvoidantigen expression level
Core Design Contradiction:
ProductivityVSQuantity of substance

Solution Approach 1:

The patent applies codon optimization to the GP antigen gene sequence, changing the nucleotide composition to match the codon usage preferences of mammalian cells. This parameter change significantly enhances the translation efficiency and protein expression level of the GP antigen produced by the AdMax-packaged adenovirus, resolving the contradiction between production efficiency and antigen expression level.

Inventive Principle:
Principle #35Parameter changes

Data Source

PatentEP3342865B1Ebola virus disease vaccine taking human replication deficient adenovirus as vector
Publication Date: 2023.09.20 INST OF BIOENG ACAD OF MILITARY MEDICAL SCI OF THE CHINESE
  • EP3342865B1 patent drawingFigure 1A~1B
  • EP3342865B1 patent drawingFigure 2~3
  • EP3342865B1 patent drawingFigure 4~6

AI summary

Provided are an Ebola virus envelope glycoprotein (that is GP protein) codon optimized nucleotide sequence, a human replication deficient recombinant adenovirus capable of expressing the nucleotide sequence, and applications in preparing a vaccine for preventing Ebola virus diseases. The nucleotide sequence takes a replication deficient 5 type adenovirus that is lack of E1 and E3 in a combined mode as a vector, HEK293 cells that integrate adenovirus E1 genes serve as a packaging cell line, and carried protective antigenic genes are codon optimized Zaire type Ebola virus Makona strain envelope glycoprotein genes. After the envelope glycoprotein genes are optimized by codon, the expression level in transfection cells is obviously improved.