Recombinant Factor C Protein for Thermostable Endotoxin Detection
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
The existing reliance on limulus amebocyte lysate for endotoxin detection poses sustainability concerns and the need for thermostable and environmentally friendly alternatives is unmet, while current recombinant Factor C solutions are hindered by patent exclusivity and supply stability issues.
Innovation Solution
Development of a recombinant Factor C protein with enhanced thermostability through amino acid substitutions, achieving high sequence identity to SEQ ID NO: 3, and a method for its production and use in endotoxin detection.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If limulus amebocyte lysate is used for endotoxin detection, then endotoxin measurement can be performed, but sustainability and supply stability are compromised
Solution Approach 1:
The patent creates a recombinant Factor C protein that copies the essential functional properties of native Factor C from limulus amebocyte lysate. By expressing the Factor C gene in recombinant systems (E. coli, insect cells, or mammalian cells), the patent produces a sustainable alternative that maintains endotoxin detection capability while eliminating dependence on finite horseshoe crab resources.
Solution Approach 2:
The patent optimizes various parameters of Factor C production including expression systems, purification conditions, and protein formulation to achieve both functional equivalence to native Factor C and improved stability. The recombinant approach allows precise control over production parameters, enabling scalable and consistent supply.
2Reliability
If conventional Factor C is used, then endotoxin detection is possible, but thermostability is insufficient
Solution Approach 1:
The patent employs protein engineering strategies to modify Factor C's amino acid sequence, introducing stabilizing mutations that enhance thermostability without compromising endotoxin binding affinity or catalytic activity. These parameter changes at the molecular level allow the protein to maintain functionality under elevated temperature conditions.
Solution Approach 2:
The patent may incorporate Factor C with stabilizing additives or formulate it in composite systems that protect against thermal degradation. The recombinant production system allows for co-expression of stabilizing proteins or incorporation of protective molecules that enhance overall thermostability.
3Stability of the object's composition
If recombinant Factor C is developed as an alternative, then supply stability improves, but manufacturing complexity increases
Solution Approach 1:
The patent divides the Factor C production process into distinct modular stages: gene synthesis/cloning, expression in selected host systems, purification through affinity and chromatography steps, and formulation. This segmentation allows each module to be optimized independently and facilitates scalable production while maintaining quality control.
Solution Approach 2:
The patent develops a universal recombinant production platform that can be applied to produce Factor C in multiple host systems (bacterial, insect, or mammalian cells). This multi-functional approach provides flexibility to select the most appropriate system based on specific requirements, reducing overall manufacturing complexity through standardization.
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
Provides a stable and reliable alternative to limulus amebocyte lysate for endotoxin detection, ensuring consistent supply and improved thermostability, addressing sustainability and supply chain concerns.
Implementation Method 1
Endotoxin convers Factor C as an enzyme precursor into activated Factor C. The activated Factor C functions as a hydrolase to convert Factor B as an enzyme precursor into activated Factor B.
Implementation Method 2
a recombinant Factor C protein... used to measure endotoxin
Data Source
Figure 1
AI summary
The present disclosure provides a means that enables detection of endotoxin. The present disclosure also provides a reagent for detecting endotoxin with thermostability. The present disclosure provides a recombinant Factor C protein having a given amino acid sequence, a polynucleotide encoding the same, a reagent for detecting endotoxin comprising the same, and a method for producing the same. The present disclosure also provides a recombinant Factor C protein having given amino acid substitution, a polynucleotide encoding the same, a reagent for detecting endotoxin comprising the same, and a method for producing the same.