Khib-Specific Affinity Reagents for Precise Protein Modification Detection
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Solution Overview
Problem
There is a need for reagents and methods to detect post-translational modifications of histones or nonhistone proteins, particularly for detecting lysine 2-hydroxyisobutyrylation, which is relevant to various diseases and disorders.
Innovation Solution
The use of peptides comprising 2-hydroxyisobutyrylated lysine to develop reagents and methods for detecting protein lysine 2-hydroxyisobutyrylation, including the production of lysine 2-hydroxyisobutyrylation specific affinity reagents such as proteins or antibodies, and the use of these reagents to form binding complexes for detection.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional detection methods are used for post-translational modifications, then general detection capability is maintained, but specific detection of lysine 2-hydroxyisobutyrylation cannot be achieved
Solution Approach 1:
The invention segments the detection approach by developing modification-specific antibodies (such as anti-K2ohibu antibodies) that target individual post-translational modification types. This allows precise detection of specific modifications like lysine 2-hydroxyisobutyrylation while maintaining the ability to detect other modifications through separate specific antibodies, thus resolving the contradiction between detection specificity and versatility.
Solution Approach 2:
The invention introduces peptide probes and affinity reagents as intermediaries that specifically recognize and bind to lysine 2-hydroxyisobutyrylation sites. These intermediaries enable indirect but highly specific detection of the target modification, allowing the system to achieve both high specificity for K2ohibu and maintain adaptability through the use of different intermediary reagents for different modifications.
2Reliability
If peptides with surrounding sequences are used for reagent production, then binding specificity is improved, but manufacturing complexity increases
Solution Approach 1:
The invention extracts the critical binding determinant (the 2-hydroxyisobutyrylated lysine residue) from the complex peptide context. By focusing on the modified lysine site and using it as the primary target for antibody generation and affinity reagent design, the invention achieves high binding specificity while simplifying the manufacturing process, as the key functional element can be produced and characterized independently of extensive surrounding sequences.
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 the specific detection of 2-hydroxyisobutyrylated lysine in proteins, providing insights into gene transcriptional activity and potential biomarkers for diseases.
Implementation Method 1
An isolated lysine 2-hydroxyisobutyrylation specific affinity reagent is also provided. It is capable of binding specifically to a peptide comprising a 2-hydroxyisobutyrylated lysine.
Data Source
AI summary
The invention provides an isolated peptide comprising a lysine 2-hydroxyisobutyrylation site, a lysine 2-hydroxyisobutyrylation specific affinity reagent that specifically binds to the peptide, and a method for detecting protein lysine 2-hydroxyisobutyrylation in a sample using the reagent.