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In this study, a new monolithic stationary phase with hyperbranched nanostructures was chemically synthesized. For this, monoliths with a support matrix of poly (glycidyl methacrylate\u2010<jats:italic>co<\/jats:italic>\u2010ethylene dimethacrylate) and ethylenediamine chemistry were modified with third\u2010generation dendrons with butyl\u2010end groups. The new monolith was analyzed by infrared spectroscopy, confirming the dendron with butyl ligands and exhibited low mass transfer resistance as observed by breakthrough frontal analysis. This support was able to separate mono\u2010PEG ribonuclease A from the PEGylation mixture, indicated by a single band (\u223c30\u00a0kDa) in the electrophoretic analysis. Moreover, the separation of mono\u2010PEGylated positional isomers was probably observed, as the protein with \u223c30\u00a0kDa was found in two separate peaks. Interestingly, the dendronized monolith allowed the separation of the reaction mixture into individual PEGylated species when using high ammonium sulfate concentrations (2\u00a0M). A correlation between the PEGylation degree and the strength of the hydrophobic interactions on the monolith was observed. This chromatographic approach combines the natural branched architecture of dendrons and the higher capabilities of the monoliths enhancing the hydrophobic surface area, and therefore the interaction between the PEGylated proteins and ligands. Thus, the novel support represents a novel platform for the purification of PEGylated from non\u2010PEGylated proteins with biotechnological applications.<\/jats:p>","DOI":"10.1002\/elps.202300098","type":"journal-article","created":{"date-parts":[[2023,8,22]],"date-time":"2023-08-22T02:16:44Z","timestamp":1692670604000},"page":"1989-1999","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["A monolithic stationary phase with dendritic nanostructures for the separation of PEGylated proteins"],"prefix":"10.1002","volume":"44","author":[{"given":"Beatriz E.","family":"De los Santos\u2010Gonz\u00e1lez","sequence":"first","affiliation":[{"name":"School of Engineering and Science Tecnologico de Monterrey Campus Monterrey, Monterrey, Nuevo  Le\u00f3n M\u00e9xico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Celeste C.","family":"Ibarra\u2010Herrera","sequence":"additional","affiliation":[{"name":"School of Engineering and Science Tecnologico de Monterrey Campus Monterrey, Monterrey, Nuevo  Le\u00f3n M\u00e9xico"},{"name":"School of Engineering and Science Tecnologico de Monterrey Campus Puebla  Puebla M\u00e9xico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jes\u00fas A.","family":"Valencia\u2010Gallegos","sequence":"additional","affiliation":[{"name":"School of Engineering and Science Tecnologico de Monterrey Campus Monterrey, Monterrey, Nuevo  Le\u00f3n M\u00e9xico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marco A.","family":"Mata\u2010G\u00f3mez","sequence":"additional","affiliation":[{"name":"School of Engineering and Science Tecnologico de Monterrey Campus Monterrey, Monterrey, Nuevo  Le\u00f3n M\u00e9xico"},{"name":"School of Engineering and Science Tecnologico de Monterrey Campus Puebla  Puebla 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