Utilizing a library of synthetic affinity ligands for the enrichment, depletion and one‐step purification of leech proteins

Abstract

Although the concept of affinity purification using synthetic ligands had been utilized for many years, there are few articles related to this research area, and they focus only on the affinity purification of specific protein by a defined library of synthetic ligands. This study presents the design and construction of a 700-member library of synthetic ligands in detail. We selected 297 ligand columns from a 700-member library of synthetic ligands to screen leech protein extract. Of the 297, 154 columns had an enrichment effect, 83 columns had a depletion effect, 36 columns had a one-step purification effect, and 58 columns had a one-step purification via flowthrough effect. The experimental results achieved by this large library of affinity ligands provide solid convincing data for the theory that affinity chromatography could be used for the enrichment of proteins that are present in low abundance, the depletion of high abundance proteins, and one-step purification of special proteins.

The Improvement of LC-MS/MS Proteomic Detection with Biomimetic Affinity Fractionation

“…It has been verified that many specific synthetic affinity ligands could be designed to purify different single proteins in our lab and other labs (Li et al, 1998;Teng et al, 1999;Gupta and Lowe, 2004;Melissis et al, 2007;Xin et al, 2007;Dong et al, 2008a). Our lab has constructed affinity ligand library (thousands of ligands), and different ligands showed different absorbance effect to proteins such as purification, depletion of high abundance proteins, and enrichment of low abundance proteins (Dong et al, 2008b), which gives a imaginable application prospect in proteomics. In this Chapter, from biomimetic affinity ligands library in our lab, we first screened out some affinity ligands showing large absorption differences in band distribution, then, selected the affinity ligands having medium absorption ability; and finally a cascade and a tandem composition of these affinity ligands were contructed and applied in prefractionation of complex tissue proteins.…”

Design and synthesis of affinity ligands and relation of their structure with adsorption of proteins

“…The affinity ligand library was constructed according to the methods developed previously [1,11,14]. Briefly, Sepharose 4B was first activated with epichlorohydrin, and then with saturated ammonia.…”

A novel fractionation method prior to MS-based proteomics analysis using cascade biomimetic affinity chromatography

“…Our lab has constructed a large affinity ligand library composed of thousands of ligands with different protein absorbance effects. This library enables us to simplify complex tissue samples through depletion of high-abundance proteins and enrichment of low-abundance proteins [21], giving this library a prospect of broad application in proteomics.…”