Zenger, OkanBaydemir Peşint, Gözde2025-01-062025-01-0620211064-374510.1007/978-1-0716-1629-1_62-s2.0-85113650789https://doi.org/10.1007/978-1-0716-1629-1_6https://hdl.handle.net/20.500.14669/1420A sensitive, rapid, and cost-effective method for quantitative analysis of proteins (e.g., detection, purification, depletion) for a wide variety of purposes is required in a number of areas, such as immunodiagnostics and biotechnology. Double-layer imprinting technique, which is carried out via polymerization of polymer solution with higher monomer concentration, covering and filling the supermacroporous structure of a pre-synthesized cryogel column with a lower monomer concentration, thus improving the surface area and adsorption capacity of final product, is a brand new approach for the application of cryogels in molecular imprinting technology. Within the scope of this chapter, BSA is selected as a model protein for the application of double-layer imprinting protocol. In this chapter, synthesis of double-layer BSA-imprinted and non-imprinted cryogel columns (BSA-DLIP and DLNIP, respectively) are described. In addition, characterization of synthesized columns and BSA depletion studies from aqueous solutions are described in detail, as well as selectivity of BSA-DLIPs for BSA, against competitors. © 2021, The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.eninfo:eu-repo/semantics/closedAccessAdvanced adsorption capacityBovine serum albuminCryogelDepletionDouble-layer imprinted polymerHigh-abundant proteinsMolecular imprinting techniqueBook Chapter8334410660Q4712359