Mutated Blocking Reagent Reduces Non-Specific Binding
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Solution Overview
Problem
Conventional antibodies used in immunoassays often exhibit non-specific binding to cell surfaces, leading to undesirable background signals that compromise the accuracy of antigen detection due to their 'sticky' nature, which existing blocking reagents may not adequately address.
Innovation Solution
Development of blocking reagents with specific mutations in their complementarity determining regions (CDRs) that reduce non-specific binding to cell surfaces, allowing for the use of antibodies that specifically bind to targeted antigens while minimizing background noise in assays.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional antibodies are used to detect antigens, then antigen detection capability is provided, but non-specific binding to cell surfaces occurs leading to high background signals
Solution Approach 1:
The patent applies parameter changes by mutating the complementarity determining regions (CDRs) of the antibody to alter its binding properties. Specifically, amino acid substitutions are made in the CDRs to reduce non-specific binding affinity while preserving specific antigen recognition, thereby changing the binding parameters of the antibody to resolve the contradiction between detection capability and background noise
Solution Approach 2:
The patent applies local quality by making targeted mutations only in the CDR regions of the antibody while leaving the rest of the antibody structure intact. This localized modification allows the antibody to maintain its overall structure and specific binding capabilities while only altering the regions responsible for non-specific binding interactions with cell surfaces
2Object-generated harmful factors
If blocking reagents are used to reduce non-specific binding, then background signal is reduced, but cross reactivity with assay components may occur disrupting specific interactions
Solution Approach 1:
The patent applies the taking out principle by extracting or removing the harmful non-specific binding property from the antibody through targeted mutations in the CDR regions. By selectively mutating specific amino acids responsible for non-specific binding while preserving the core antigen-binding capability, the harmful aspect is separated from the useful aspect of antibody function
Solution Approach 2:
The patent applies inversion by creating an antibody variant that has reduced non-specific binding properties while maintaining specific binding capability. Instead of trying to enhance specific binding to overcome non-specific binding, the approach is inverted: non-specific binding is actively reduced through mutations, allowing specific binding to dominate without requiring excessive enhancement
Data Source
AI summary
Blocking reagent compositions, as well as methods of making and using the same, are provided. Aspects of the composition include a blocking reagent that binds to the cell surface but does not specifically bind to a targeted first cell surface antigen. Also provided are system and kits that include the compositions.