Nucleic Acid Array Protein Capture for Multiplex Antibody Screening
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Solution Overview
Problem
Current antibody screening methods, such as tissue microarrays, suffer from low throughput and accuracy issues due to species specificity, requiring multiple tissue sections and prone to false positives/negatives and background interference, limiting their effectiveness in high-throughput screening.
Innovation Solution
A nucleic acid array-based detection composition using probes and protein-nucleic acid conjugates for high-throughput antibody screening, enabling simultaneous detection of multiple antibodies on the same tissue section, with spatial localization and sequencing to avoid background interference.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If tissue microarrays are used for antibody screening, then high-throughput screening of tissue types is enabled, but the throughput is limited to only 1-4 antibody types due to species specificity
Solution Approach 1:
The patent introduces nucleic acid tags as intermediaries between antibodies and detection systems. Each antibody is conjugated with a unique nucleic acid tag, allowing multiple antibodies to be simultaneously detected through nucleic acid-based methods rather than traditional chromogenic or fluorescent detection, thereby enabling ultra-high-throughput screening of dozens of antibody types
Solution Approach 2:
The patent replaces the traditional mechanical/optical detection system (chromogenic reagents and fluorescence visualization) with a nucleic acid-based detection system. This substitution eliminates the limitation of detecting only 1-4 antibody types simultaneously and enables parallel detection of numerous antibodies through nucleic acid sequencing and analysis
2Reliability
If multiple tissue sections are used for negative and positive control stainings, then control accuracy is improved, but significant gaps between sections lead to differences in cellular composition and antigen type
Solution Approach 1:
The patent merges the detection of multiple antibodies, including negative and positive controls, onto a single tissue section. By using nucleic acid tags conjugated to each antibody and detecting them simultaneously through nucleic acid-based methods, the system eliminates the need for multiple separate tissue sections, ensuring that all controls and test antibodies are applied to the identical cellular composition and antigen landscape
Solution Approach 2:
The patent creates a universal detection platform where a single tissue section can serve multiple functions: detecting test antibodies, negative controls, and positive controls simultaneously. The nucleic acid-based detection system allows all these different antibody types to be identified in parallel on the same section, making the section universally applicable for all screening purposes
3Measurement precision
If chromogenic reagents or fluorescence are used to visualize antibody locations, then positive results can be identified, but background staining and autofluorescence affect identification accuracy
Solution Approach 1:
The patent replaces the chromogenic and fluorescent detection systems with a nucleic acid-based detection system. Instead of using chromogenic reagents that produce background staining or fluorescent markers that suffer from autofluorescence interference, the system uses nucleic acid tags that are detected through specific hybridization and sequencing methods, completely eliminating background staining issues and significantly improving signal-to-noise ratio
Solution Approach 2:
The patent extracts the detection signal from the traditional optical domain (chromogenic and fluorescent signals) and transfers it to the nucleic acid domain. By conjugating nucleic acid tags to antibodies and detecting these tags through hybridization and sequencing, the system removes the harmful background staining and autofluorescence that are inherent to optical detection methods
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 ultra-high-throughput screening of dozens of antibodies with reduced false positives/negatives by using nucleic acid arrays to capture and sequence antibodies, providing accurate spatial information and avoiding background staining issues.
Implementation Method 1
The nucleic acid array probe comprises a capture sequence and the capture sequence is used for capturing the protein-nucleic acid conjugate
Implementation Method 2
the amplification adaptor B sequence is used for amplifying
Implementation Method 3
the excision region is used for releasing an amplification product from a carrier
Implementation Method 4
the spatial localization sequence is used for spatially localizing the nucleic acid array probe
Implementation Method 5
the UMI sequence is used for quantifying and removing a duplicate sequence introduced by amplification
Data Source
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AI summary
The present invention relates to the field of biotechnology, and in particular to a detection composition and a protein capture detection method. Provided is a detection composition, comprising a nucleic acid array probe and a protein-nucleic acid conjugate, wherein the nucleic acid array probe comprises a capture sequence, and the capture sequence is used for capturing the protein-nucleic acid conjugate. Simultaneous detection of ultra-high multiplex antibodies can be performed on a same tissue slice. Because spatial reduction of the antibody position depends on sequencing technology, the background staining problem caused by traditional immunohistochemistry and immunofluorescence color developing agents can be solved. In addition, negative control, positive control, and detection of an antibody under test can be carried out on the same tissue slice, a detection result can be determined more accurately, and a false-negative or false-positive result caused by traditional staining technology is eliminated.