High-Throughput Protein Glycoprofiling With Nested Signal-Amplifying Beads
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Solution Overview
Problem
Existing methods for determining the glycoprofile of proteins lack sufficient sensitivity, selectivity, and speed, particularly for detecting cancer-specific markers.
Innovation Solution
A method involving the use of first beads with antibodies specific to proteins, followed by further beads with labels and lectins to form antibody-protein-lectin complexes, allowing for signal amplification and low-level detection of glycan structures.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional antibody-lectin assays are used for glycoprofiling, then the method is relatively simple, but the sensitivity and detection limit are insufficient
Solution Approach 1:
The patent combines multiple functions into a single bead structure. The first bead contains both the capture antibody for protein enrichment and the lectin for glycan detection, eliminating the need for separate enrichment and detection steps. This merging increases sensitivity while managing complexity through functional integration.
Solution Approach 2:
The patent employs a nested bead structure where a first bead (containing capture antibody) is combined with a second bead (containing lectin and label). The second bead is introduced to the complex formed by the first bead and target protein, creating a nested antibody-protein-lectin complex. This nesting enables signal amplification and improved detection sensitivity.
2Productivity
If conventional glycoprofiling methods are used, then the procedure is straightforward, but the analysis time is too long
Solution Approach 1:
The patent performs preliminary enrichment of the target protein using the capture antibody on the first bead before the actual glycan detection step. This preliminary action concentrates the target analyte, reducing the time needed for subsequent detection steps and overall analysis time while maintaining straightforward methodology.
Solution Approach 2:
By merging the enrichment function (capture antibody) and detection function (lectin with label) into a single integrated bead system, the patent eliminates multiple sequential steps required in conventional methods. This combination accelerates the overall analysis while keeping the procedure relatively simple.
3Measurement precision
If conventional assays are used for cancer biomarker detection, then the method is simple, but the selectivity for cancer-specific markers is insufficient
Solution Approach 1:
The patent applies local quality by using specific lectins that recognize particular glycan structures associated with cancer markers. Each lectin is selected for its ability to bind to specific aberrant glycans on cancer-related proteins, providing high selectivity for cancer-specific markers while maintaining a relatively simple detection approach.
Solution Approach 2:
The lectin acts as an intermediary between the capture antibody and the detection label. It specifically recognizes and binds to cancer-associated glycan structures on the captured protein, providing selectivity for cancer markers. This intermediary function enhances detection specificity without requiring complex multi-step procedures.
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
Enhances sensitivity and selectivity in glycoprofiling, particularly for cancer biomarkers, with reduced analysis time.
Implementation Method 1
contacting a sample comprising said protein with first beads having coupled thereto an antibody directed against said protein, to form an antibody-protein complex
Implementation Method 2
contacting said antibody-protein complex with one or more further beads, each further bead having coupled thereto (i) a label which amplifies a signal being generated and (ii) a lectin, to form an antibody-protein-lectin complex
Implementation Method 3
a label which amplifies a signal being generated
Data Source
AI summary
The present invention discloses a method of determining the glycoprofile of a protein, comprising (a) contacting a sample comprising said protein with first beads having coupled thereto an antibody directed against said protein, to form an antibody-protein complex, (b) contacting said antibody-protein complex with one or more further beads, each further bead having coupled thereto (i) a label which amplifies a signal being generated and (ii) a lectin, to form an antibody-protein-lectin complex; and (c) determining the glycoprofile of said protein. Further disclosed are methods for diagnosing cancer, autoimmune diseases and inflammatory diseases as well as kits for performing the methods disclosed herein.