Antigen-Specific ELISPOT Detection for Infection Diagnosis
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Conventional Elispot methods for diagnosing infections and diseases face challenges such as poor signal-to-background ratios, increased risk of false-negative and false-positive results, and non-specific stimulation leading to unspecific antibody secretion and apoptosis, which affect the accuracy of antibody-secreting cell detection.
Innovation Solution
A method involving specific stimulation of antibody-secreting cells with a large number of differently labeled antigens, where the antigens are used for both stimulation and detection, allowing antigen-specific antibodies to bind to immobilized anti-Ig antibodies, thereby improving signal-to-background ratios and reducing non-specific binding.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If non-specific stimulation is used to stimulate antibody-secreting cells, then cell activation occurs, but signal-to-background ratio deteriorates due to spontaneous apoptosis and non-specific antibody secretion
Solution Approach 1:
The patent segments the stimulation process by using separate specific antigens for each antibody-secreting cell population rather than a single non-specific stimulus. This allows targeted activation of relevant cells while avoiding activation of unrelated cells that would contribute to background noise and false signals.
Solution Approach 2:
The patent introduces specifically labeled antigens as intermediaries that mediate between the detection system and antibody-secreting cells. These antigens serve as precise mediators that only activate cells producing antibodies against the specific antigen, thereby eliminating non-specific activation and improving signal-to-background ratio.
2Reliability
If non-specific stimulation is applied, then general cell activation occurs, but false-negative and false-positive results increase
Solution Approach 1:
The patent divides the detection process into specific antigen-targeted segments, where each antigen population is detected separately using its specific labeled antigen. This segmentation prevents cross-reactivity and non-specific activation, thereby reducing false-negative and false-positive results while maintaining reliable cell activation detection.
Solution Approach 2:
Specifically labeled antigens act as precise intermediaries that ensure only the intended antibody-secreting cells are activated and detected. This intermediary approach eliminates cross-reactivity and non-specific binding, thereby improving detection accuracy and reducing false results.
3Reliability
If antibodies are secreted during non-specific stimulation, then cell response is generated, but binding sites on carrier surface are competed for, reducing sensitivity
Solution Approach 1:
The patent segments the antibody secretion detection by using antigen-specific stimulation for each target. This ensures that antibodies secreted during stimulation are specific to the labeled antigen and can bind to the corresponding immobilized antibodies on the carrier surface without being competed for by non-specific antibodies, thereby maintaining high detection sensitivity.
Solution Approach 2:
Specifically labeled antigens serve as precise intermediaries that induce secretion of only the relevant antibodies. This eliminates competition from non-specific antibodies for binding sites on the carrier surface, thereby preserving detection sensitivity while maintaining reliable cell response 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
This approach enhances the accuracy of detecting antibody-secreting cells by minimizing false results and improving specificity, allowing for better diagnosis and monitoring of infections, diseases, and vaccination responses.
Implementation Method 1
antibodies which are produced by di e stimulated antibody-secreting cells are produced, bind to the immobilized anti-Ig antibodies of the support surface and the antigens bind to antibodies specific for the antigens
Implementation Method 2
bind to the immobilized anti-Ig antibodies of the support surface
Data Source
Figure 1a
Figure 1b
Figure 1c
AI summary
The invention relates to a method for the in vitro diagnosis and/or in vitro monitoring of the therapy of an infection and/or disease, in particular an infectious disease, and/or tumor disease, and/or autoimmune disease, and/or allergy, and/or for vaccination monitoring, and/or for the detection of cross-protectiveness, wherein the method comprises the following steps: (a) providing - a sample containing antibody-secreting cells, - a carrier with a surface on which anti-Ig antibodies are immobilized, and - a plurality of different antigens from various infectious pathogens and/or various tumor antigens and/or various autoimmune antigens and/or various allergens, wherein the antigens are each labeled differently, (b) bringing the carrier surface into contact with the plurality of different antigens and the sample under conditions suitable for the absorption of antibodies,which are produced by antibody-secreting cells, bind to the immobilized anti-Ig antibodies on the carrier surface and bind the antigens to antibodies specific for the antigens, and (c) detect labeled antigens captured on the carrier surface, thereby detecting the presence or absence of antibody-secreting cells specific for the antigens contained in the sample.