Human serum albumin-imprinted polymers with high capacity and selectivity for abundant protein depletion. (May 2021)

• Record Type: Journal Article
• Authors: Lu, Wenjun; Wang, Sha; Liu, Rui; Guan, Ying; Zhang, Yongjun
• Is Part Of: Acta biomaterialia. Volume 126(2021)
• Page Start: 249
• Page End: 258
• Publication Date: 2021-05
• Subjects: Protein-imprinted polymers -- Proteomic analysis -- Human serum albumin -- pH response -- Helix-coil transition; Biomedical materials -- Periodicals; 610.28
• DOI: 10.1016/j.actbio.2021.03.010
• Languages: English
• ISSNs: 1742-7061

Abstract

Abstract: Depletion of human serum albumin (HSA), the most abundant protein in human plasma, from serum/plasma is a prerequisite before their proteomic analysis. Molecularly imprinted polymers (MIPs) using HSA as a template have been designed for this purpose, but suffer from a low sorption capacity and low selectivity. Here, a new HSA-imprinted polymer was synthesized using N-isopropylacrylamide (NIPAM) as the main monomer; acrylamide (AAm), methacrylic acid (MAA), and dimethylaminoethyl methacrylate (DMAEMA) as functional monomers; and oligoglutamic acid-based peptide crosslinker (PC) as a crosslinker at pH 5.5. When pH is adjusted to 7.4, the peptide chains in the polymer change from a helical conformation to an extended coil conformation, and the polymer swells. Consequently, the template protein is removed completely. When pH is adjusted back to 5.5, the peptide chains fold back precisely to the helical conformation. Both the size and shape of the imprint cavities are restored. Therefore, the polymer rebinds the template protein selectively. Highest imprinting factor (IF) was observed at pH 5.5 at which the polymer was synthesized. The IF increases with the increasing number of glutamic acid residues in the PCs because of their increased degree of helicity at pH 5.5. No improvement in imprinting effect was observed when using a peptide crosslinker containing both L- and D-glutamic acid residues and hence incapable of folding into α-helix, further confirming the key role of the pH-induced helix-coil transition of the peptide chains. The MIP synthesized here presents a much higher affinity to HSA than the nontemplate proteins. It could be used repeatedly without evident decrease in sorption capacity. Because of the mild eluting conditions, the secondary structure of the extracted HSA protein remains unchanged. Finally, the MIP was used to deplete HSA from human serum. Because of its high sorption capacity and high selectivity, HSA was depleted completely and selectively. Statement of significance: A new molecularly imprinted polymer (MIP) using human serum albumin (HSA) as a template was synthesized using N-isopropylacrylamide (NIPAM) as the main monomer; acrylamide (AAm), methacrylic acid (MAA), and dimethylaminoethyl methacrylate (DMAEMA) as functional monomers; and oligoglutamic acid-based peptide crosslinker as a crosslinker. Because of the reversible and precise pH-induced helix-coil transition of the peptide chains, the template protein was removed facilely and completely under mild conditions. Simultaneously, a significant improvement in imprinting efficiency was obtained. The sorption capacity was as high as 648.05 mg/g and the imprinting factor was 7.9. Because of its high selectivity and high binding capacity, the MIP synthesized here is highly promising for the depletion of HSA, the most abundant protein in serum, which is a prerequisite for its proteomic analysis. For the first time, complete and selective depletion of HSA from human serum was achieved using a protein-imprinted polymer. Graphical abstract: Image, graphical abstract