Zenger, OkanPesint, Gozde Baydemir2025-01-062025-01-0620221359-51131873-329810.1016/j.procbio.2022.03.0182-s2.0-85128982976https://doi.org/10.1016/j.procbio.2022.03.018https://hdl.handle.net/20.500.14669/2578In this study, bilayer-imprinted cryogels were synthesized for depletion of human serum albumin (HSA) from human serum. For this purpose, a monolayer HSA-imprinted cryogel with 8% monomer concentration was synthesized and second imprinting process was performed using a polymeric solution with 16% monomer concentration in presence of HSA, in the supermacroporous structure of first layer. Thereby, bilayer HSAimprinted cryogel column (HSA-BLIP) with enhanced surface area was obtained. Bilayer non-imprinted polymer (BL-NIP) and monolayer HSA-imprinted polymers with monomer concentrations of 8% and 16% (HSAMIP(8%) and HSA-MIP16%, respectively) were synthesized as the control group. Synthesized columns were characterized by swelling tests, scanning electron microscopy (SEM), multipoint Brunauer-Emmett-Teller (BET) surface area measurement and micro computed tomography (Micro-CT) analysis. Surface areas of HSA-BLIP, BL NIP, HSA-MIP8% and HSA-MIP(16% )were found to be 39 m(2)/g, 37 m(2)/g, 21 m(2)/g and 12 m(2)/g, respectively. Maximum HSA adsorption was calculated as 106.8 mg/g for HSA-BLIP and 5.14 mg/g for BL-NIP. Selectivity studies were conducted in presence of competitor molecules, human hemoglobin (Hb) and immunoglobulin G (IgG). Finally, selective depletion of HSA directly from human serum was studied. In addition, it was demonstrated that HSA-BLIPs can be used up to 10 times without significant decrease in selective adsorption capacity.eninfo:eu-repo/semantics/closedAccessHuman serum albuminCryogelsBilayer-imprintingMolecular ImprintingProtein DepletionArticle100Q190117WOS:000806790000001Q2