Affinity on Demand: A One-Pot Method for Synthesis and Sample Enrichment Using TentaGel-Functionalized Resins

Area of Science:

• Biochemistry
• Proteomics
• Analytical Chemistry

Background:

• Protein glycation, forming Amadori products, is implicated in diabetes complications.
• Analyzing low-abundance glycated peptides in biological samples presents significant proteomic challenges.
• Selective analyte enrichment is essential for overcoming detection limitations.

Purpose of the Study:

• To develop a novel, efficient method for synthesizing functionalized resins for glycated peptide enrichment.
• To evaluate the affinity and selectivity of the synthesized resins for glycated peptides.
• To demonstrate the applicability of the resin in analyzing complex biological samples.

Main Methods:

• A one-pot synthesis method for creating phenylboronic acid-functionalized resins with tunable properties.
• Application of the highest-affinity resin (4PhB-3Lys-TGR) for selective enrichment of glycated peptides from albumin hydrolysate, patient serum, and milk.
• Bioinformatic analysis to assess the coverage of protein sequences identified from enriched glycated peptides.

Main Results:

• Successfully synthesized functionalized resins with high affinity for glycated peptides.
• Demonstrated selective enrichment of glycated peptides from various complex biological matrices.
• Bioinformatic analysis confirmed high protein sequence coverage, validating the enrichment method's effectiveness.

Conclusions:

• A novel, fast, simple, and cost-effective one-pot method for synthesizing resins for selective glycated peptide enrichment has been developed.
• The developed resin and enrichment method are effective for proteomic analysis of glycated peptides in complex samples.
• The approach holds potential for affinity-based isolation of other compound classes, including Amadori products and carbonyl compounds.