Immunoassay Anti-Poly-Histidine Antibody Interference Removal
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Solution Overview
Problem
Immunoassays often produce false-negative and false-positive results due to the presence of anti-poly-histidine antibodies, which interfere with the detection of target antibodies in biological samples, particularly in assays using recombinantly produced proteins with poly-histidine tags, leading to inaccurate disease diagnosis and tissue compatibility testing.
Innovation Solution
A method involving the use of poly-histidine polymers to block or remove anti-poly-histidine antibodies from biological samples before exposure to target antigens, using affinity purification techniques and kits with solid-phase substrates immobilized with histidine polymers to prevent interference and enhance assay accuracy.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of manufacture
If poly-histidine tagged recombinant proteins are used as antigens in immunoassays, then the ease of manufacture and purification of antigens is improved, but false-positive and false-negative results occur due to anti-poly-histidine antibody interference
Solution Approach 1:
The patent removes the harmful poly-histidine tag from the recombinant antigen used in the immunoassay. By expressing the antigen without the C-terminal poly-histidine sequence that causes auto-antibody interference, the assay maintains manufacturing simplicity while eliminating false-positive and false-negative results caused by anti-poly-histidine antibody reactions
Solution Approach 2:
The patent modifies only the C-terminal region of the recombinant antigen by removing the poly-histidine tag while preserving the rest of the antigen structure. This localized modification eliminates the harmful epitope recognized by auto-antibodies while maintaining the antigen's ability to bind specific antibodies of interest
2Quantity of substance
If anti-poly-histidine antibodies are present in biological samples, then the quantity of antibodies detected increases, but the measurement precision decreases due to false-positive and false-negative results
Solution Approach 1:
The patent converts the harmful effect of anti-poly-histidine antibodies into a beneficial outcome by using these antibodies as a marker to identify and select suitable test subjects. Subjects with high levels of anti-poly-histidine antibodies are identified and excluded from testing, ensuring that only subjects with low or absent anti-poly-histidine antibodies undergo the immunoassay, thereby eliminating false-positive and false-negative results
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
Significantly reduces false-positive and false-negative results by specifically removing anti-poly-histidine antibodies, thereby improving the sensitivity and specificity of immunoassays for detecting target antibodies, ensuring accurate disease diagnosis and tissue compatibility testing.
Implementation Method 1
A method involving the use of poly-histidine polymers to block or remove anti-poly-histidine antibodies from biological samples before exposure to target antigens, using affinity purification techniques
Data Source
AI summary
Described herein are materials and methods for improved immunoassays using synthetic antigens in which samples are treated to bind antibodies specific to histidine polymer antigens.