Effective protein adsorption by solid matrices from complex biological samples has attracted attention for broad application in biomedical field. Immobilization of calixarenes to solid supports is an essential process for their application in protein separation and purification. Silica is the most widely used support material in calixarene immobilization. With high concentration of polymer microspheres as templates, the large pore mesoporous silica microspheres with controllable, uniform size and structure were successfully synthesized and the resulting large pore mesoporous silica microspheres were modified with water-soluble tetrasulfonate calix[4]arene of unique hollow cavity-shaped structure. The tetrasulfonate calix[4]arene modified large pore mesoporous silica microspheres (SCLX4@LPMS) were characterized by diverse analytical techniques and their protein adsorption performance were also investigated. The obtained SCLX4@LPMS gave rise to an adsorption efficiency of >90% for cytochrome c and lysozyme within a wide pH range of 3.0-10.0 and possessed remarkably high adsorption capacity of cytochrome c (363.64 mg g-1) and lysozyme (166.11 mg g-1). The retained cytochrome c and lysozyme can be readily eluted by using phosphate buffer solution containing NaCl as a stripping reagent with the recoveries of 81% and 86% after 5 times enrichment, respectively. The SCLX4@LPMS microspheres have been applied for the selective adsorption of proteins in real samples and had the application potential in protein adsorption, drug delivery, biosensors, and other biomedical fields.
Keywords: Calixarene; Cytochrome c; Lysozyme; Mesoporous silica microspheres; Protein adsorption.
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