DNA Aptamer Selective Detection of Monomeric CRP
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Solution Overview
Problem
Current methods for detecting C-reactive protein (CRP) in body fluids, such as ELISA, are unable to selectively quantify monomeric CRP (mCRP) due to cross-reactivity with pentameric CRP (pCRP), and existing aptamer-based methods are not specific for mCRP.
Innovation Solution
Development of DNA aptamers that specifically bind to mCRP, allowing for its selective detection and quantification in body fluid samples without binding to pCRP, using sequences like 5′-tg gcg ggt tgt gaa ggg tgg agt atg gtc gtg ttg gtt-3′, and employing methods like ELISA, SPR, or capillary electrophoresis for analysis.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If commercial antibodies (e.g., CRP-8 antibody) are used for CRP detection, then CRP can be detected in body fluids, but cross-reactivity with pCRP prevents selective quantification of mCRP
Solution Approach 1:
The invention changes the detection parameter from antibody-based recognition to aptamer-based recognition. The aptamer sequence 5'-kg rss ksk krs srd drk dkk rkd rwr vdv kkg dkr gtk-3' was specifically selected to bind mCRP through its unique structural features, thereby changing the binding specificity parameter to achieve selective detection without cross-reactivity with pCRP
Solution Approach 2:
The invention uses a DNA aptamer as a molecular copy or surrogate that mimics the binding function of antibodies but with superior specificity. The aptamer serves as an alternative recognition element that copies the detection function while eliminating the cross-reactivity issue inherent in commercial antibodies
2Ease of manufacture
If aptamer-based detection methods are developed, then cost and production ease improve, but existing aptamers lack specificity for mCRP
Solution Approach 1:
The invention optimizes the aptamer sequence parameters through systematic evolution of ligands by exponential enrichment (SELEX). The specific sequence 5'-kg rss ksk krs srd drk dkk rkd rwr vdv kkg dkr gtk-3' was generated through multiple selection cycles to achieve high specificity for mCRP while maintaining the ease of in vitro production characteristic of aptamers
Solution Approach 2:
The invention replaces the need for complex animal-based antibody production systems with a purely in vitro aptamer selection system. This substitution maintains the cost-effectiveness and ease of manufacture of aptamers while achieving the required specificity through biochemical selection rather than biological immunization
3Productivity
If classical ELISA methods are used, then detection capability is established, but inability to distinguish mCRP from pCRP limits diagnostic utility
Solution Approach 1:
The invention segments the CRP detection function into two distinct detection systems: one for total CRP using conventional methods and one for selective mCRP detection using the specific aptamer. This segmentation allows simultaneous measurement of both isoforms with different detection reagents, enabling precise discrimination between mCRP and pCRP contributions to total CRP levels
Solution Approach 2:
The DNA aptamer acts as an intermediary recognition element that mediates specific binding to mCRP. The aptamer sequence 5'-kg rss ksk krs srd drk dkk rkd rwr vdv kkg dkr gtk-3' serves as a molecular bridge that selectively connects the detection system to mCRP without cross-reacting with pCRP, thereby enabling precise measurement
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 DNA aptamers provide a sensitive and specific tool for detecting and quantifying mCRP, distinguishing it from pCRP, thereby offering a reliable diagnostic platform for inflammation and other inflammatory conditions.
Implementation Method 1
DNA aptamers that specifically bind to monomeric CRP, allowing for its selective detection and quantification
Data Source
AI summary
The present invention is directed to selective detection of monomeric C-reactive protein (CRP) in a body fluid sample. In particular, the present invention provides the use of a DNA aptamer for the detection of monomeric CRP in an isolated sample of body fluid from a subject. The invention also provides DNA aptamers that specifically bind to monomeric CRP. Finally, also a detection method for monomeric CRP in a body fluid sample using a DNA aptamer and a kit for the detection of monomeric CRP in a body fluid sample are described.