Polysaccharide Support Conjugate Reagents via Reductive Amination
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
There is a need for effective conjugate reagents that can accurately determine the presence and amount of analytes in samples using specific binding assays, particularly those involving polysaccharide layers and amine-reactive functionalities, where existing methods face challenges in forming stable linkages without amide bonds.
Innovation Solution
A composition and method involving a support with an inner and outer polysaccharide layer, where the outer layer has pendant moieties with terminal amine functionalities that form linkages with specific binding pair members using a linking compound with two terminal amine groups, one reacting with the aldehyde group and the other with the amine-reactive functionality, avoiding amide bond formation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional amide bond formation is used to conjugate specific binding pair members to supports, then stable linkages are formed, but the coupling efficiency is reduced and binding affinity is compromised
Solution Approach 1:
The invention changes the chemical parameters of the linkage by using reductive amination to form C=N bonds instead of conventional amide bonds. This parameter change in bond type enables both high coupling efficiency and good binding affinity, resolving the contradiction between linkage stability and coupling efficiency
Solution Approach 2:
The invention introduces an intermediary step of forming a C=N bond through reductive amination as a mediator between the support and the specific binding pair member. This intermediary mechanism allows for efficient coupling while maintaining stable linkages, overcoming the limitations of direct amide bond formation
2Ease of manufacture
If conventional conjugation methods are used, then supports can be prepared, but accurate analyte detection is compromised due to reduced binding affinity
Solution Approach 1:
By changing the chemical parameters of the conjugation process to use reductive amination instead of conventional methods, the invention maintains ease of manufacture while significantly improving measurement precision through enhanced binding affinity and coupling efficiency
3Ease of manufacture
If amide bond linkages are formed between the outer polysaccharide layer and specific binding pair members, then conjugates can be formed, but binding affinity is reduced
Solution Approach 1:
The invention changes the fundamental parameter of bond type from amide bonds to C=N bonds through reductive amination. This parameter change maintains the ease of conjugate formation while dramatically improving binding affinity, as the C=N bond provides better preservation of the specific binding pair member's binding characteristics
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 formation of conjugates with enhanced coupling efficiency and good binding affinity, allowing for accurate analyte detection in assays, as demonstrated by the universal reagent's ability to form conjugates with various specific binding pair members, including haptens and steroid hormones.
Implementation Method 1
reacting the terminal amine functionality of the composition with the member of the specific binding pair, which comprises an amine-reactive functionality
Implementation Method 2
One of the terminal amine groups reacts with the aldehyde group of the outer polysaccharide layer to form a non-amide bond
Data Source
Figure 1
AI summary
Compositions are disclosed that include a support having on a surface thereof at least an inner polysaccharide layer and an outer polysaccharide layer. The outer polysaccharide layer has pendant moieties each comprising a terminal amine functionality. The point of linkage of the pendant moiety to the outer polysaccharide layer does not comprise an amide bond. The composition may be employed in a method of conjugating a member of a specific binding pair to a particle. The method comprises reacting the terminal amine functionality of the composition with the member of the specific binding pair, which comprises an amine-reactive functionality.