Lamprey VLR-B Fusion for Recombinant Protein Multimerization
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Solution Overview
Problem
Recombinant proteins often fail to maintain the native three-dimensional conformation and quaternary structure, leading to altered or suppressed biological function, particularly in oligomeric proteins like influenza haemagglutinin, where poor trimerization results in low immunogenicity.
Innovation Solution
Fusion of a nucleic acid sequence encoding a protein of interest with a sequence from the lamprey variable lymphocyte receptor B (VLR-B) antibody, specifically the C-terminal domain, to promote multimerization and stability of recombinant proteins, retaining biological activity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If recombinant proteins are produced without fusion to multimerization sequences, then the production process is simpler, but the proteins fail to maintain native quaternary structure and biological activity
Solution Approach 1:
The patent uses a multimerization sequence (such as the foldon domain from T4 phage or other known multimerization domains) as an intermediary element that is fused to the recombinant protein. This intermediary sequence mediates the formation of multimeric structures, enabling the recombinant protein to maintain its native quaternary structure and biological activity without requiring complex production conditions.
Solution Approach 2:
The patent creates composite protein structures by fusing the recombinant protein sequence with a multimerization sequence. This results in a chimeric protein that combines the functional domain of the native protein with the structural multimerization capability of the fused sequence, thereby achieving stable quaternary structure formation.
2Reliability
If recombinant proteins are produced without fusion to multimerization sequences, then the expression system is simpler, but the immunogenicity is reduced due to poor trimerization
Solution Approach 1:
The multimerization sequence acts as an intermediary that facilitates proper oligomeric assembly of the recombinant protein. This intermediary element ensures that the protein forms the correct multimeric structure (e.g., trimers for influenza HA), which is essential for maintaining immunogenicity and eliciting effective immune responses.
Solution Approach 2:
The patent modifies the molecular parameters of the recombinant protein by fusing it with a multimerization sequence. This changes the protein's self-association properties, enabling it to form stable multimers with appropriate stoichiometry and geometry, thereby improving immunogenicity without requiring complex expression system modifications.
3Stability of the object's composition
If fusion to lamprey VLR-B sequence is used, then multimerization and stability are improved, but the protein sequence becomes more complex
Solution Approach 1:
The C-terminal domain of lamprey VLR-B serves as a multimerization mediator that confers stable oligomeric structure to the fused recombinant protein. This intermediary domain promotes the formation of stable multimeric complexes while maintaining the functional integrity of the native protein portion.
Solution Approach 2:
The lamprey VLR-B multimerization sequence exhibits universal applicability, as it can be fused to various different recombinant protein sequences to promote multimerization. This multi-functional element provides a general solution for stabilizing diverse recombinant proteins without requiring protein-specific optimization.
Data Source
AI summary
The present invention relates to polymerized recombinant proteins, to recombinant nucleic acids coding for the polymerized recombinant proteins, to expression cassettes comprising the recombinant nucleic acids, to host cells transformed by the expression cassettes and to a method for multimerizing a recombinant protein. The polymerized proteins of the invention may be used in pharmaceutical or immunogenic compositions. In particular, the recombinant proteins may be antigens, antibodies or scaffolds. In particular, the polymerized recombinant protein may be an influenza haemagglutinin.