LEDGF-Modified Mammalian Cells for ANA Detection

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Solution Overview

Problem

Current methods for detecting anti-nuclear antibodies (ANAs) using HEp-2 cells are prone to false results and complex interpretation due to the presence of anti-DFS70 antibodies, which are common in healthy individuals and can complicate the diagnosis of autoimmune diseases.

Innovation Solution

Modified mammalian cells that either do not express or express reduced levels of the LEDGF protein are used in diagnostic assays, allowing for the differentiation of antibodies binding to LEDGF protein by comparing binding patterns between LEDGF- and LEDGF+ cells, thereby reducing false positives and improving test accuracy.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Adaptability or versatility

If HEp-2 cells are used as substrate for ANA testing, then the test can detect a broad range of autoantigens (100-150 relevant auto-antigens), but false positive results increase due to anti-DFS70 antibodies present in healthy individuals

Engineering Contradiction:

Improvedetection range of autoantigensVSAvoidtest accuracy

Core Design Contradiction:

Adaptability or versatilityVSReliability

Solution Approach 1:

The patent segments the detection process by using two different cell substrates: HEp-2 cells for detecting ANA patterns and LEDGF-negative cells for detecting anti-DFS70 antibodies. This segmentation allows the test to maintain broad detection capability while eliminating false positives by separating the detection of different antibody types into distinct testing steps.

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

The patent extracts and removes the source of false positives by using LEDGF-negative cells that have had the LEDGF/DFS70 protein gene deleted or knocked down. This extraction of the problematic antigen from the testing substrate eliminates interference from anti-DFS70 antibodies while preserving the ability to detect other ANA patterns using HEp-2 cells.

Inventive Principle:

Principle #2Taking out (Extraction)

2Measurement precision

If HEp-2 cells are used for ANA detection, then comprehensive autoantigen detection is achieved, but test interpretation complexity increases due to multiple patterns and false results

Engineering Contradiction:

Improvedetection capabilityVSAvoidtest interpretation complexity

Core Design Contradiction:

Measurement precisionVSDevice complexity

Solution Approach 1:

The patent divides the complex interpretation task into two simpler, separate interpretations: first interpreting ANA patterns from HEp-2 cell staining, then interpreting anti-DFS70 antibody presence from LEDGF-negative cell staining. This segmentation reduces interpretation complexity by eliminating the need to differentiate between true positive and false positive DFS70 patterns within a single test.

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

The patent introduces LEDGF-negative cells as an intermediary testing substrate that acts as a control mechanism. By comparing results between HEp-2 and LEDGF-negative cells, the system provides a clear mediator for distinguishing true ANA positivity from false positives, simplifying the overall interpretation process.

Inventive Principle:

Principle #24Intermediary (Mediator)

3Reliability

If LEDGF-negative cells are used to detect anti-DFS70 antibodies, then false positives are reduced, but the ability to detect other ANA patterns may be limited

Engineering Contradiction:

Improvereduction of false positivesVSAvoiddetection of diverse ANA patterns

Core Design Contradiction:

ReliabilityVSAdaptability or versatility

Solution Approach 1:

The patent merges the results from two separate cell-based assays: HEp-2 cell staining for comprehensive ANA pattern detection and LEDGF-negative cell staining for anti-DFS70 antibody detection. By combining these two tests in a sequential or parallel manner, the system achieves both high reliability in reducing false positives and maintained versatility in detecting diverse ANA patterns.

Inventive Principle:

Principle #5Merging (Combining)

Solution Approach 2:

The patent creates a multi-functional diagnostic approach where LEDGF-negative cells serve the dual function of detecting anti-DFS70 antibodies specifically while also serving as a control to confirm true ANA positivity. This universal application of the modified cells enhances both test reliability and interpretative value without limiting detection capabilities.

Inventive Principle:

Principle #6Universality (Multi-functionality)

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 ANA detection by reducing the DFS pattern interference, providing a more reliable diagnostic tool for autoimmune disorders by distinguishing between healthy and diseased individuals.

Implementation Method 1

ANA detection using IIF assay can reveal a multitude of patterns such as homogeneous, fine granular, coarse granular, nucleolar, centromere, nuclear dots, pleomorphic, mitochondrial and a variety of cytoskeletal patterns

Methodology Applied

Scientific EffectIndirect immunofluorescence: Fluorescence

Data Source

PatentEP3916007A1Improved Anti-nuclear antibody detection and diagnostics for systemic and non-systemic autoimmune disorders

Publication Date: 2021.12.01 IMMCO DIAGNOSTICS

Figure 1A1~1B2

AI summary

Provided are compositions that contain mammalian cells for use in detecting antibodies. The mammalian cells are modified such that they do not contain LEDGF protein. The mammalian cells are immobilized on a solid substrate. The compositions can also contain mammalian cells that contain the LEDGF protein. Methods for using the cell compositions in diagnostic approaches are included, as are kits for performing diagnostic tests.