3D cryogel composites as adsorbent for isolation of protein and small molecules

Abstract

A green and promising sample pretreatment method was successfully established, which efficiently isolated proteins and small molecules in human serum. This method was achieved based on the multifunctional polymer, cryogel, as a solid phase extraction (SPE) monolith easily equipped in a syringe. The cryogel (pDC/GO-DE) was composed of diallyldimethyl ammonium chloride (DC) and 2-hydroxyethyl methacrylate (HE), which was further modified with graphene oxide (GO) and N-diethylethanamine hydrobromide (DE). Various proteins, including bovine serum albumin (BSA), lysozyme (Lys), γ-globulins, immunoglobulin G (IgG), transferrin, small molecules (ribavirin, adenosine, ofloxacin, estriol, rutin, amoxicillin, ibuprofen, 1-methyl-3-phenyl-propylamine, and benzylamine) and their mixtures were successively studied as model analytes to evaluate the new material and demonstrate the isolation mechanism, which was mainly dependent on mixed-mode ion-exchange and the hybrid hydrophobicity-hydrophilicity property of pDC/GO-DE cryogel. Moreover, the three-dimensional macroporous structure contributed to the underlying size-selective isolation. When 10 times diluted human serum was used as the sample, more than 95% of proteins were adsorbed within 10 min under physiological conditions, and the interference matrix in serum was also efficiently reduced. After recycling three times, the extraction ratio of proteins in human serum was still higher than 90%. When four small molecules (camptothecin, ribavirin, 1-methyl-3-phenylpropylamine and ofloxacin) were added to blank human serum, their recoveries were within 65.6-81.8%, and were comparable to those obtained by protein precipitation method (63.7-83.2%).

Keywords: Adsorbent; Cryogel composite; Human serum; Pretreatment; Protein precipitation; Sample; Self-assembly.