Synthesis of Novel Hybrid Adsorption-Permeation Adsorbents for Effective Separation of Peptides and Small Proteins

Abstract

The main objective of this work was to synthesize new adsorbents for the isolation and separation of peptides and small proteins from mixtures of larger biomolecules found in biological fluids. This was done by constructing hybrid gels with both adsorption and size-exclusion chromatography (AdSEC) characteristics on the same matrix. The ligands were reacted onto the surface to create adsorption sites. The ligands functionalized onto surface were chelating agents including iminodiacetic acid (IDA), dipicolylamine (DPA), carboxymethylated ethylenediamine (CEDA), tripicolylamine (TPA) and amino acids including arginine, lysine, glycine, aspartic acid, phenylalanine, asparagine, and histidine and a couple derivatives of ethylenediamine (EDA). The hindrance permeating compound used in all these AdSEC gels was the polymer methyl polyethylene glycol amine (m-PEG-NH2). Proof of concept was performed with synthesized IDA-PEG-m AdSEC gels to perform separations of amino acids from bovine serum albumin (BSA) solutions. Studies were also performed with DPA-PEG and with carboxymethylated ethylenediamine (CEDA)-PEG derivatives. Tests with DPA and CEDA included sample solutions containing different peptides. Functionalized gel analysis showed that reaction times of both the specific adsorption ligand and the permeation polymer PEG onto the matrix is the most important parameter to produce optimal adsorption-permeation chromatographic matrices for effective separation of larger and small molecular weight biomolecules. The new gels synthesized in this work offer an effective tool to isolate and separate target biomolecules from complex biological mixtures such as blood, urine, sweat and tears.