Lactylated Lysine Affinity Reagents for 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 linked to various diseases and disorders, particularly those associated with the Warburg effect and its non-metabolic functions in physiology and disease.
Innovation Solution
Development of affinity reagents that specifically bind to lactylated lysines in peptides, including antibodies, and methods for detecting and isolating these reagents using peptide libraries or immunization, along with kits for lactylated lysine detection and isolation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional detection methods are used for post-translational modifications, then general protein detection is possible, but specific detection of lactylated lysine is not achievable
Solution Approach 1:
The patent uses a biotinylated peptide as an intermediary to isolate and enrich lactylated lysine-containing peptides from complex protein samples. This intermediary enables specific capture of the target modification through streptavidin-biotin interaction, allowing precise detection of lactylated lysine that cannot be achieved by conventional methods
2Measurement precision
If affinity reagents are developed for specific binding, then detection precision is improved, but the complexity of reagent development increases
Solution Approach 1:
The patent employs a self-service approach where the lactylated peptide itself serves as the basis for generating the affinity reagent. By using the target peptide to immunize animals or select phage display libraries, the system automatically generates antibodies or binding proteins that specifically recognize the lactylated lysine modification, simplifying the overall development process
3Reliability
If protein libraries are screened for binding specificity, then reliable affinity reagents are obtained, but the time and resources required increase
Solution Approach 1:
The patent performs preliminary enrichment of lactylated peptides using biotinylated peptide capture before subjecting the enriched samples to library screening or immunization. This preliminary action concentrates the target modification and removes background interference, making subsequent affinity reagent development more efficient and reliable while reducing overall time requirements
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 detection and quantification of lactylated lysines in proteins, providing insights into lactate's role in pathophysiological conditions such as infection and cancer, and understanding the epigenetic regulation by histone lactylation.
Implementation Method 1
an affinity reagent that binds specifically a lactylated lysine in a peptide
Data Source
AI summary
The invention provides an isolated peptide comprising a lactylated lysine and a specific affinity reagent that specifically binds to a lactylated lysine in a peptide. Also provided are a method for detecting a lactylated lysine in a protein or a fragment thereof using the affinity reagent and a method for isolating the affinity reagent.