TSH Receptor Autoantibody Assay Using cAMP Biosensor Cells
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Solution Overview
Problem
Existing methods for measuring TSH receptor autoantibody activity are time-consuming and require steps like medium adaptation of cells or pretreatment of blood samples, and they lack accuracy in determining thyroid diseases like Basedow's disease.
Innovation Solution
A method and kit that utilize mammal cells expressing both a cAMP biosensor and TSH receptor to measure autoantibody activity by incubating with a blood sample, measuring activation levels, and comparing them to controls, without requiring special apparatus or lengthy pretreatments.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional bioassay methods are used to measure TSAb activity, then measurement accuracy is improved, but measurement time increases to 44 hours or longer
Solution Approach 1:
The patent applies preliminary action by pre-adapting cells to the measurement medium before actual measurement, and pre-removing endogenous cAMP from serum samples. This preparation is done in advance so that the actual measurement can be completed rapidly without compromising accuracy. The cells are adapted to the measurement conditions beforehand, and serum is pretreated to remove interfering substances, enabling fast measurement while maintaining reliability.
Solution Approach 2:
The patent extracts and removes endogenous cAMP from serum samples using activated carbon treatment before measurement. This extraction of the interfering substance (endogenous cAMP) eliminates the need for long incubation periods and allows for rapid accurate measurement. By taking out the harmful factor (endogenous cAMP that interferes with measurement), the measurement can be completed quickly without compromising precision.
2Loss of time
If serum samples are not pretreated to remove endogenous cAMP, then measurement time is reduced, but measurement accuracy deteriorates due to interference
Solution Approach 1:
The patent converts the harmful effect of endogenous cAMP in serum into a beneficial situation by using activated carbon to selectively remove it. The endogenous cAMP, which originally interfered with measurement accuracy, is transformed from a harmful factor into a removed substance, allowing rapid measurement without accuracy loss. This converts the problem of endogenous cAMP interference into a solution where its removal enables fast measurement.
Solution Approach 2:
The patent introduces activated carbon as an intermediary substance to remove endogenous cAMP from serum samples. This intermediary agent facilitates the separation and removal of interfering substances without requiring complex procedures or long incubation times. The activated carbon acts as a mediator that selectively binds and removes endogenous cAMP, enabling rapid accurate measurement.
3Reliability
If cells are adapted to measurement medium before measurement, then measurement reliability is improved, but total time required increases
Solution Approach 1:
The patent applies preliminary action by performing cell adaptation to the measurement medium in advance, before the actual measurement process. This preliminary adaptation ensures that cells are properly acclimated to measurement conditions, improving reliability. By doing this preparation beforehand, the actual measurement can be completed rapidly without compromising the reliability that would otherwise require extended adaptation periods.
4Loss of time
If existing rapid methods are used, then measurement time is reduced to 4-6 hours, but accuracy in determining thyroid disease severity is insufficient
Solution Approach 1:
The patent replaces the conventional cAMP measurement system with a luminescence-based detection system. Instead of measuring cAMP directly through enzymatic reactions that require long incubation periods, the patent uses a luminescence assay that provides rapid and sensitive detection. This substitution of the measurement mechanism enables both speed and accuracy, allowing for rapid determination of TSAb activity levels that correlate with disease severity.
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 method allows rapid and accurate determination of TSH receptor autoantibody activity within 3 hours, distinguishing between stimulatory and inhibitory activities, and identifying thyroid disease severity, reducing false negatives and enabling appropriate treatments.
Implementation Method 1
measuring an activation level of the cAMP biosensor
Data Source
AI summary
Provided herein is a method and kit that accurately measure autoantibody activity against a TSH receptor without pretreating a blood sample. The autoantibody activity against a TSH receptor in a blood sample is measured by using a kit for measurement of autoantibody activity against a TSH receptor in a mammal cell, comprising a mammal cell expressing both a cAMP biosensor and the TSH receptor, and a substrate capable of visualizing and/or quantifying the cAMP biosensor as constituents, and performing the steps of: (a) incubating a mammal cell expressing both a cAMP biosensor and the TSH receptor in the presence of a blood sample collected from a test subject; (b) measuring an activation level of the cAMP biosensor after the step (a); and (c) comparing the activation level measured in the step (b) with an activation level in a control to calculate the autoantibody activity.