Lateral Flow FXIII Detection via Glutamine Substrate
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Solution Overview
Problem
Current methods for detecting blood coagulation factor XIII (FXIII) are inadequate for rapid, accurate determination, especially in acute situations, due to interference from blood components and the need for laboratory-based tests that are slow and laborious.
Innovation Solution
A point-of-care compatible test principle using a lateral flow device with an immobilized glutamine donor substrate, where FXIII binds to the substrate and an amine donor, allowing for detection through spectral changes, which is not influenced by interfering components like serum-albumin and fibrinogen.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional coagulation tests (thromboplastin time, aPTT) are used to detect FXIII deficiency, then the test can be performed in standard laboratory settings, but these tests are not suitable for detecting FXIII deficiency and require complex sample preparation and long processing times
Solution Approach 1:
The test system is segmented into distinct functional zones on a lateral flow strip: a sample application zone, a reaction zone containing immobilized glutamine donor substrate, and a detection zone. This spatial segmentation enables rapid, point-of-care testing by separating sample processing from detection, eliminating the need for complex laboratory procedures and reducing processing time while maintaining detection accuracy for FXIII activity
Solution Approach 2:
An immobilized glutamine donor substrate serves as an intermediary on the lateral flow strip that specifically interacts with FXIII. When FXIII in the blood sample contacts this substrate, it catalyzes the binding of an amine donor to the glutamine donor, producing a detectable signal. This intermediary enables direct, rapid detection of FXIII activity without requiring complex laboratory equipment or lengthy processing
2Productivity
If photometric or fluorimetric determination is performed in whole blood, then direct measurement is possible, but strong light absorption by hemoglobin and interference from blood components prevent accurate detection
Solution Approach 1:
The harmful interfering components (hemoglobin, serum albumin, fibrinogen) are effectively removed from the detection process by using a lateral flow approach where only the relevant FXIII-enzyme complex is retained on the strip. The blood sample is applied to the strip, and through capillary action and selective binding, interfering components flow through without interacting with the detection system, while FXIII is specifically captured and measured, enabling rapid testing without interference
Solution Approach 2:
The immobilized glutamine donor substrate acts as a selective intermediary that binds specifically to FXIII while excluding other blood components. This specific interaction ensures that only FXIII activity is measured, eliminating interference from hemoglobin's light absorption and other blood constituents, thereby enabling rapid and accurate productivity
3Loss of time
If rapid separation of erythrocytes using glass fiber mats is performed, then plasma can be obtained quickly, but soluble blood constituents remain in the plasma and interfere with FXIII detection
Solution Approach 1:
The immobilized glutamine donor substrate on the lateral flow strip serves as a selective intermediary that specifically captures FXIII from the complex blood matrix. Even when soluble blood constituents like serum albumin and fibrinogen remain in the plasma, they do not interfere with the specific FXIII-substrate interaction. The substrate selectively binds FXIII through the transglutaminase reaction, enabling rapid sample preparation without requiring complete purification
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
Enables rapid and accurate determination of FXIII activity in human blood samples with minimal sample preparation and interference, providing a reliable method for on-site assessment during emergencies.
Implementation Method 1
Activated FXIII as a transglutaminase links proteins by forming an isopeptide bond between a γ-glutamine residue and the ε-amino group in an exposed lysine side chain. Ammonia is released during this reaction.
Implementation Method 2
The number of the bond between amine donor and FXIII substrate is determined by the temporal change of the spectroscopic signal. The decrease of NADH is measured via the absorbance at 340 nm and it is directly proportional to the FXIII activity.
Implementation Method 3
The decrease of NADH is measured via the absorbance at 340 nm
Implementation Method 4
The bond takes place to an immobilized glutamine donor substrate. The number of the bond between amine donor and FXIII substrate is determined by the temporal change of the spectroscopic signal.
Data Source
AI summary
A method and a device for the detection of transglutaminases, in particularly of blood coagulation factor XIII (FXIII). The method is based on a rapid, point-of-care compatible test principle for transglutaminases and allows an exact determination of FXIII in a short time and with simple handling.