Dual-Peptide Target Binding Moieties for Sensitive Antibody Profiling
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Solution Overview
Problem
Existing methods for interrogating antibody repertoires, such as sequencing and binding approaches, face limitations in generating large enough and sensitive libraries of molecules for antibody profiling, particularly due to challenges in preparing peptide arrays with high sensitivity and binding affinity.
Innovation Solution
A composition comprising a plurality of polynucleotide-barcoded target binding moieties with linked first and second peptide sequences separated by a spacer, allowing simultaneous binding to an antibody's antigen binding domain, and a nucleic acid sequence encoding these peptides, enabling a large and sensitive library of target-binding molecules.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional peptide arrays are used for antibody profiling, then the library can be prepared, but the sensitivity is low due to low binding affinity
Solution Approach 1:
The invention divides the binding unit into multiple peptide sequences (first and second peptide sequences) that can simultaneously bind to different antigen binding domains of the antibody. This segmentation allows each peptide to contribute to the overall binding affinity while the combined structure achieves high sensitivity detection
Solution Approach 2:
The invention creates a composite structure combining multiple peptide sequences with different functionalities - some peptides provide binding affinity while others provide solubility or stability. This composite approach resolves the contradiction by integrating materials with complementary properties to achieve both high binding affinity and high sensitivity
2Quantity of substance
If a large library of peptides is prepared for antibody profiling, then more targets can be examined, but the preparation becomes time-consuming and difficult
Solution Approach 1:
The invention designs a universal library structure where all peptides share common features (such as solubility tags or structural frameworks) while varying only in the binding region. This allows a large library size to be maintained while simplifying preparation procedures, as the common features can be introduced through standardized methods
Solution Approach 2:
The invention performs preliminary actions by pre-introducing solubility-enhancing sequences or structural elements into all library members during synthesis. This preliminary preparation ensures that all peptides in the large library are soluble and stable without requiring individual optimization, thus maintaining both large library size and high preparation efficiency
Data Source
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AI summary
Provided herein are compositions and methods to identify a binding element (e.g., peptide, peptoid, or protein) that can be bound by an immunoreceptor (e.g., antibody). The binding element can be provided in a target binding unit comprising two binding elements separated by a spacer such that the two binding elements simultaneously bind to a single molecule comprising an antigen binding domain of an antibody. The present disclosure provides various strategies to construct the spacer. The identified binding elements can be further used to manufacture an array which can be used to profile antibodies obtained from a blood sample.