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

## TL;DR

This paper introduces a new one-pot method to create resins that selectively enrich glycated peptides, improving their analysis in complex biological samples.

## Contribution

A novel one-pot synthesis method for functionalized resins with high affinity for glycated peptides is presented.

## Key findings

- The highest-affinity resin (4PhB-3Lys-TGR) effectively enriches glycated peptides from complex samples.
- The method was successfully applied to milk, serum, and glycated albumin hydrolyzate.
- Bioinformatics analysis confirmed high coverage of protein sequences identified via glycated peptides.

## Abstract

Protein glycation
is a nonenzymatic reaction that results in the
formation of early glycation products, commonly referred to as Amadori
products, which play an important role in diabetes complications.
In proteomic research, the analysis of glycated peptides is very challenging
due to the low amount of analyte in a biological sample. One of the
methods to overcome this is selective enrichment of the sample in
the desired analyte. A method for synthesizing functionalized resins
with phenylboronic acids has been developed, which allows for the
incorporation of different linkers and a variable number of phenylboronic
acid moieties, as well as the use of any solid support. Furthermore,
the resins are prepared for use in sample enrichment following the
completion of the synthesis process and demonstrate a high affinity
for glycated peptides. The highest-affinity resin (4PhB-3Lys-TGR)
was applied to artificially glycated albumin hydrolyzate and patient
serum, and, in addition, it was used in conjunction with a biological
sample (i.e., milk) for the selective enrichment of glycated peptides.
The bioinformatics analysis provided results that confirmed the high
coverage of protein sequences identified in the complex samples based
on glycated peptides. This paper presents a novel, fast, simple, and
cost-effective one-pot method for the synthesis of functionalized
resins, along with a selective method for the enrichment of samples
with glycated peptides. We believe that the presented approach is
general and, with necessary modifications, could be applied for affinity-based
isolation not only of Amadori products but also of carbonyl compounds,
thiols, compounds with chelating properties, and others.