Crosslinking Reagent for Bioconjugation for Use in Crosslinking Proteomics, in Particular Crosslinking Mass Spectrometry Analysis

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This patent describes a novel crosslinking reagent specifically designed for use in crosslinking proteomics, particularly in mass spectrometry analysis. The reagent features terminal protein-reactive groups and a central biotin-derivative (such as desthiobiotin) that enables reversible binding to streptavidin supports, allowing for the efficient enrichment and subsequent release of crosslinked peptide pairs under mild conditions. This provides a significant improvement over traditional biotin-based systems, which suffer from extremely strong, non-reversible binding to streptavidin.

Use CasesContent extracted from patent full text and abstract with AI.

• Identification and analysis of protein-protein interactions in complex biological samples using crosslinking mass spectrometry.
• Structural studies of proteins within living cells, organelles, or cellular lysates.
• Preparation of enriched crosslinked peptide samples for proteomic research and database matching.
• Development of commercial kits for crosslinking-based enrichment and mass spectrometry workflows in research and clinical laboratories.
• Improved workflows for affinity purification and isolation of protein complexes, particularly where reversible binding is required.

BenefitsContent extracted from patent full text and abstract with AI.

• Reversible binding and easy elution of crosslinked peptides from streptavidin supports, eliminating the need for harsh conditions that may damage samples.
• Increased efficiency and yield of crosslinked peptide enrichment, facilitating more accurate and comprehensive protein structure and interaction analysis.
• Compatibility with mild, MS-friendly buffer systems, preserving sample integrity and downstream mass spectrometry performance.
• Simplified and rapid synthesis of the reagent, making it suitable for practical laboratory and commercial applications.
• Reduced hydrophobicity and improved water solubility compared to traditional biotinylated crosslinkers, decreasing risk of aggregation and increasing reagent versatility in diverse research settings.

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Patent Abstract

The present invention relates to a crosslinking reagent for bioconjugation for use in crosslinking proteomics, in particular crosslinking mass spectrometry analysis, a method for obtaining the same and the use thereof for enriching peptide pairs.