Label-free detection microarray for novel peptide ligands screening base on MS-SPRi combination

Area of Science:

• Biomedical Engineering
• Analytical Chemistry
• Molecular Biology

Background:

• Peptide ligands with high affinity and specificity are valuable in biomedicine.
• Traditional peptide screening methods are time-consuming and labor-intensive.
• Integrated analytical approaches are needed for efficient peptide screening.

Purpose of the Study:

• To develop and demonstrate a label-free detection microarray system for one-bead-one-compound (OBOC) peptide screening.
• To facilitate the efficient screening of affinity peptides towards specific targets, such as AD protein.
• To provide a high-throughput and universal solution for ligand discovery.

Main Methods:

• A microwell array chip with two identical units for one-bead-one-compound trapping.
• In situ photo-release of peptides from beads.
• Label-free detection using Surface Plasmon Resonance imaging (SPRi) and in situ single bead sequencing by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS).

Main Results:

• Successfully screened affinity peptides towards AD protein from a library of 10^4 candidates.
• Demonstrated both qualitative and quantitative screening capabilities.
• The bi-unit imprinted chip system enabled efficient and high-throughput screening.

Conclusions:

• The developed label-free detection microarray system significantly enhances peptide screening efficiency.
• This integrated approach offers a universal solution for high-throughput ligand screening.
• The method accelerates the discovery of high-affinity peptide ligands for biomedical applications.