Dimeric WFA Lectin Carrier for Sugar Chain Detection
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Solution Overview
Problem
Existing methods for detecting target substances with sugar chains face challenges in maintaining the steric structure of lectin, leading to reduced reactivity and stability, which affects sensitivity and reproducibility.
Innovation Solution
A reagent comprising a dimeric Wisteria floribunda agglutinin (WFA) immobilized on a solid phase carrier, specifically biotinylated dimeric WFA immobilized via biotin-binding protein, is used to enhance binding capacity and storage stability, ensuring high sensitivity and reproducibility in detecting sugar chains.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If lectin is immobilized on a solid phase carrier to detect sugar chains, then detection capability is improved, but the steric structure of lectin deteriorates over time leading to reduced reactivity and stability
Solution Approach 1:
The patent segments the natural tetrameric lectin into smaller dimeric units through controlled proteolytic cleavage. This segmentation maintains the essential steric structure for sugar chain binding while reducing the overall complexity and improving stability during immobilization. The dimeric lectin retains functional activity but exhibits better structural stability when immobilized on solid phase carriers.
Solution Approach 2:
The patent changes the molecular size parameter of lectin from tetrameric to dimeric form. This parameter change optimizes the balance between maintaining sufficient steric structure for detection and improving storage stability. The smaller dimeric units are less prone to structural degradation while still preserving the key binding characteristics needed for reliable detection.
2Manufacturing precision
If natural tetrameric lectin is used to maintain steric structure, then initial reactivity is preserved, but long-term storage stability deteriorates
Solution Approach 1:
By segmenting the tetrameric lectin into dimeric units, the patent achieves a compromise between preserving reactivity and improving storage stability. The dimeric structure maintains sufficient steric configuration for sugar chain recognition while being more resistant to denaturation and degradation during long-term storage, thus extending the functional lifespan of the reagent.
Solution Approach 2:
The patent creates a more stable, longer-lasting lectin reagent that reduces the need for frequent replacement. The dimeric lectin's improved storage stability means it maintains functionality over extended periods, making it a more durable and cost-effective solution compared to conventional tetrameric lectin that degrades faster and requires more frequent replacement.
3Ease of operation
If lectin is immobilized via conventional methods, then detection function is achieved, but reactivity with sugar chains decreases over time
Solution Approach 1:
The dimeric lectin structure facilitates more effective immobilization while preserving reactivity. The smaller size and modified structure of dimeric units allow for better orientation and attachment to the solid phase carrier without compromising the steric configuration needed for sugar chain binding, thus maintaining high reactivity throughout the reagent's operational life.
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 dimeric WFA reagent maintains high reactivity with sugar chains and exhibits excellent storage stability, enabling sensitive and reproducible detection of target substances, particularly those with N-acetyl-D-galactosamine residues.
Implementation Method 1
biotinylated dimeric WFA immobilized via biotin-binding protein
Implementation Method 2
lectin that binds to the sugar chain... WFA which is lectin that binds with a sugar chain of M2BP
Data Source
AI summary
The present invention relates to a method for detecting a target substance that is useful for detection of a glycoprotein and the like comprising a specific sugar chain, a reagent for detecting a target substance, a carrier and a method for manufacturing the carrier.