Secondary Amino Alcohols: Traceless Cleavable Linkers for Use in Affinity Capture and Release

Area of Science:

• Chemical Biology
• Materials Science
• Biotechnology

Background:

• Peptide capture and release are crucial for discovering novel functional materials.
• Existing methods often present a trade-off between efficient capture and facile release.

Purpose of the Study:

• To develop novel linkers for overcoming the challenge of facile peptide release after capture.
• To introduce secondary amino alcohol-based linkers for peptide discovery platforms.

Main Methods:

• Incorporation of seramox and isoseramox linkers into peptides via solid-phase peptide synthesis using reductive amination.
• Quantitative cleavage of linkers using sodium periodate (NaIO4) treatment.
• Evaluation of linker compatibility with peptide libraries and nanoLC-MS/MS identification.
• Assessment of cell compatibility and in-cell peptide capture and identification.

Main Results:

• Both seramox and isoseramox linkers were quantitatively cleaved within minutes.
• Isoseramox cleavage yielded a native peptide N-terminus.
• The isoseramox linker demonstrated broad substrate compatibility, enabling identification of all sequences in a synthetic peptide library via nanoLC-MS/MS.
• Linkers proved cell-compatible, facilitating capture and identification of cell-penetrating peptides after cellular uptake.

Conclusions:

• Secondary amino alcohol-based linkers offer a solution for efficient peptide capture and facile release.
• These linkers are versatile tools for peptide discovery, compatible with various applications including in-cell analysis.
• The developed linkers show promise for advancing peptide-discovery platforms and the development of novel functional materials.