Recombinant Adenovirus Terminal Sequence Homogeneity

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Solution Overview

Problem

There is a need for recombinant adenovirus batches with improved homogeneity, particularly in the 5' terminal nucleotide sequences, to ensure safety and efficacy in large-scale production for clinical use, as existing methods may result in heterogeneous batches due to variations in the terminal sequences.

Innovation Solution

The introduction of a specific 5' terminal nucleotide sequence CTATCTAT (nucleotides 1-8) in recombinant adenovirus genomes, achieved through molecular cloning and plaque purification, ensures that essentially all adenovirus particles in a batch have the same terminal sequence, enhancing homogeneity and replication efficiency.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Quantity of substance

If conventional adenovirus production methods are used, then large-scale production can be achieved, but batch homogeneity deteriorates due to variations in terminal sequences

Engineering Contradiction:

Improveproduction scaleVSAvoidbatch homogeneity

Core Design Contradiction:

Quantity of substanceVSManufacturing precision

Solution Approach 1:

The patent applies parameter changes by modifying the terminal 8 nucleotides of the ITR sequence from the conventional CATCATCA to CTATCTAT. This specific sequence modification stabilizes the terminal ends during passaging, preventing heterogeneity while allowing large-scale production to proceed without compromising batch consistency.

Inventive Principle:

Principle #35Parameter changes

2Manufacturing precision

If molecular cloning and plaque purification are used to ensure homogeneity, then batch homogeneity improves, but production complexity increases

Engineering Contradiction:

Improvebatch homogeneityVSAvoidproduction process complexity

Core Design Contradiction:

Manufacturing precisionVSDevice complexity

Solution Approach 1:

The patent implements preliminary action by pre-modifying the terminal sequence to CTATCTAT before production. This upfront modification eliminates the need for repeated molecular cloning and plaque purification steps during production, as the stabilized sequence prevents heterogeneity from developing during passaging, thereby reducing overall process complexity.

Inventive Principle:

Principle #10Preliminary action

3Productivity

If terminal sequence variations occur during passaging, then replication efficiency may improve in some cases, but batch reliability deteriorates

Engineering Contradiction:

Improvereplication efficiencyVSAvoidbatch consistency

Core Design Contradiction:

ProductivityVSReliability

Solution Approach 1:

The patent applies local quality by specifically modifying only the terminal 8 nucleotides of the ITR sequence while leaving the rest of the genome unchanged. This localized modification stabilizes the terminal ends to prevent heterogeneity, ensuring batch reliability, while the canonical core region (nucleotides 9-18) remains intact to maintain replication efficiency.

Inventive Principle:

Principle #3Local quality

Data Source

PatentUS8932607B2Batches of recombinant adenovirus with altered terminal ends

Publication Date: 2015.01.13 JANSSEN VACCINES & PREVENTION BV

AI summary

Described is a composition comprising a plurality of recombinant adenovirus particles, being a recombinant human adenovirus of serotype 5, 26, 34, 35, 48, 49 or 50, or a recombinant simian adenovirus, characterized in that the genomes of essentially all adenovirus particles in the composition comprise as the 5′ terminal nucleotides the nucleotide sequence: CTATCTAT (nucleotides 1-8 of SEQ ID NO:7). Also described are methods to produce such compositions.