Recombinant PCV-2 Capsid Protein Production and Quantification
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Solution Overview
Problem
Current methods for producing and quantifying the capsid protein of Porcine circovirus 2 (PCV-2) face challenges such as low expression levels in E. coli due to rare codons and the lack of post-translational modifications in prokaryotic systems, and existing vaccines require laborious techniques for detection and quantification.
Innovation Solution
The production of recombinant capsid protein and virus-like particles (VLPs) in E. coli, using bacterial systems that overexpress rare codons, combined with the use of aluminum hydroxide as an adjuvant, and a capture ELISA kit for quantification, which allows for efficient antigen recovery and quantification.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If recombinant capsid protein is produced in E. coli, then production cost is reduced and production speed is increased, but expression level is low due to rare codons and lack of post-translational modifications
Solution Approach 1:
The patent changes the biological parameters of the E. coli system by introducing bacterial strains that overexpress rare codons (codon optimization) and modifies expression conditions (temperature, induction timing) to maximize capsid protein yield while maintaining the advantages of prokaryotic expression systems
Solution Approach 2:
The patent creates virus-like particles (VLPs) that replicate the structural and immunogenic properties of native viral capsids through recombinant protein self-assembly, producing functional copies without requiring complex eukaryotic post-translational modification systems
2Measurement precision
If traditional detection methods are used for vaccine antigens, then detection can be performed, but the process is laborious and time-consuming
Solution Approach 1:
The patent employs ELISA-based detection methods that utilize colorimetric changes as readouts for antigen quantification, enabling rapid visual or spectrophotometric detection of vaccine antigen levels without laborious procedures
Solution Approach 2:
The patent replaces manual, mechanical detection procedures with automated immunoassay systems that use antibody-antigen binding reactions and signal amplification to achieve precise quantification rapidly
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This approach enables the rapid production of tens to hundreds of milligrams of antigen in hours, with self-assembly of VLPs similar to eukaryotic systems, and precise quantification of PCV-2 antigens, enhancing vaccine efficiency and reducing production costs.
Implementation Method 1
with self-assembly of VLPs similar to eukaryotic systems
Implementation Method 2
a capture ELISA kit for quantification, which allows for efficient antigen recovery and quantification
Data Source
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AI summary
The present invention relates to the preparation of the recombinant antigen of the viral capsid of Porcine circovirus 2 (PCV-2) and modifications thereof, upon expression in a prokaryotic system, purification in the monomer form, recovery of virus-like particles (VLPs) and their use in vaccine formulations, diagnostic kits and a system for quantifying in vaccine lots of the PCV-2 antigen by means of a capture ELISA assay. The antigens and vaccine formulations can be used in animal's immunization in programs for combatting PCV-2-associated diseases in conventional swine breeding systems, and represent alternatives to the commercially available vaccines. The ELISA kit can be used for testing the quality of commercial and/or experimental vaccines against PCV-2.