Chimeric Fusion Protein for Reduced Diagnostic Interference
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Solution Overview
Problem
Current methods for producing chaperone-like proteins for biotechnological and diagnostic applications often result in interferences due to non-human amino acid sequences, leading to high background noise or incorrect test results, and lack an effective folding helper with both high catalytic and chaperone activities.
Innovation Solution
A chimeric fusion protein is created by fusing the polypeptide binding segment of a non-human chaperone protein with sequences from an FK506 binding protein or an FK506-binding-protein-like domain, specifically with human peptidyl-prolyl-cis/trans isomerase, to produce a humanized PPIase chaperone with superior folding helper activities.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If non-human chaperone proteins are used for protein folding, then folding helper activity is provided, but interference and background noise occur in diagnostic tests
Solution Approach 1:
The invention applies local quality by creating a chimeric protein where only specific domains retain chaperone function while other domains are humanized. The non-human chaperone domains (providing folding activity) are strategically positioned and limited to specific regions, while human domains minimize interference in diagnostic applications.
Solution Approach 2:
The invention uses composite materials by constructing a chimeric fusion protein that combines non-human chaperone domains with human protein domains. This composite structure integrates the beneficial folding helper activity of non-human proteins while incorporating human sequences to reduce immunogenicity and background noise in diagnostic tests.
2Object-affected harmful factors
If human FKBP sequences are used, then interference in diagnostic tests is minimized, but catalytic and chaperone activities are insufficient
Solution Approach 1:
The invention merges the advantages of both human and non-human protein sequences by creating a chimeric fusion protein. The human FKBP domains minimize diagnostic interference while the integrated non-human chaperone domains provide enhanced catalytic and chaperone activities, achieving superior overall performance.
Solution Approach 2:
The chimeric fusion protein achieves multi-functionality by combining domains with different functions: human FKBP domains for reduced interference and non-human chaperone domains for enhanced folding and catalytic activities. This universal design allows the single protein to fulfill multiple requirements simultaneously.
3Power
If complete non-human chaperone proteins are used, then high catalytic efficiency is achieved, but immunogenicity and diagnostic interference increase
Solution Approach 1:
The invention applies segmentation by dividing the chaperone protein into distinct functional domains and selectively humanizing certain regions. This segmentation allows retention of catalytically active non-human domains while replacing immunogenic portions with human sequences, reducing overall immunogenicity while maintaining enzymatic function.
Solution Approach 2:
Local quality is applied by making the protein partially human rather than completely human or non-human. Specific regions are humanized to reduce immunogenicity and diagnostic interference, while other regions maintain non-human sequences to preserve high catalytic efficiency.
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
The resulting chimeric fusion proteins exhibit enhanced catalytic efficiencies and chaperone properties, minimizing interference in diagnostic tests and pharmaceutical applications while maintaining native-like folded protein structures.
Implementation Method 1
nucleotide sequences coding for an FK506 binding protein (FKBP) or an FK506-binding-protein-like domain (FKBP-like domain)
Data Source
AI summary
The invention discloses the cloning, expression and uses of a chimeric fusion protein with superior chaperone and folding activities compared to the wild type chaperones. This invention relates to a chimeric fusion protein encoded by a recombinant DNA molecule containing nucleotide sequences coding for a polypeptide binding segment of a non-human chaperone protein and nucleotide sequences coding for an FK506 binding protein (FKBP) or an FK506-binding-protein-like domain (FKBP-like domain). In particular, this invention relates to a chimeric fusion protein encoded by a recombinant DNA molecule containing nucleotide sequences coding for a polypeptide binding segment of a non-human chaperone protein and nucleotide sequences coding for a human FKBP type peptidyl-prolyl-cis/trans isomerase (PPIase), methods of producing these chimeric fusion proteins and their uses as folding helpers in the production of other proteins and in the process of the production of vaccines or pharmaceuticals, and as folding helpers for performing immunoassays.