title: Advances in organic polymer-based monolithic column technology for high-resolution liquid chromatography-mass spectrometry profiling of antibodies, intact proteins, oligonucleotides, and peptides.
abstract: This review focuses on the preparation of organic polymer-based monolithic stationary phases and their application in the separation of biomolecules, including antibodies, intact proteins and protein isoforms, oligonucleotides, and protein digests. Column and material properties, and the optimization of the macropore structure towards kinetic performance are also discussed. State-of-the-art liquid chromatography-mass spectrometry biomolecule separations are reviewed and practical aspects such as ion-pairing agent selection and carryover are presented. Finally, advances in comprehensive two-dimensional LC separations using monolithic columns, in particular ion-exchange×reversed-phase and reversed-phase×reversed-phase LC separations conducted at high and low pH, are shown.
authors.affiliation: Vrije Universiteit Brussel, Department of Chemical Engineering, Pleinlaan 2, B-1050 Brussels, Belgium. Electronic address: seeltink@vub.ac.be.
authors.forename: Sebastiaan
authors.initials: S
authors.lastname: Eeltink
authors.affiliation: Vrije Universiteit Brussel, Department of Chemical Engineering, Pleinlaan 2, B-1050 Brussels, Belgium.
authors.forename: Sam
authors.initials: S
authors.lastname: Wouters
authors.affiliation: Vrije Universiteit Brussel, Department of Chemical Engineering, Pleinlaan 2, B-1050 Brussels, Belgium.
authors.forename: José Luís
authors.initials: JL
authors.lastname: Dores-Sousa
authors.affiliation: Beijing Advanced Innovation Center for Soft Matter Science and Engineering, Beijing University of Chemical Technology, Beijing, China.
authors.forename: Frantisek
authors.initials: F
authors.lastname: Svec
mesh_terms: D000906:Antibodies / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification; D002623:Chemistry Techniques, Analytical / Q000295:instrumentation*; D002851:Chromatography, High Pressure Liquid; D013058:Mass Spectrometry; D009841:Oligonucleotides / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification; D010455:Peptides / Q000737:chemistry*; D011108:Polymers / Q000737:chemistry; D020033:Protein Isoforms / Q000032:analysis / Q000737:chemistry; D011506:Proteins / Q000032:analysis*
chemical_list: D000906:Antibodies; D009841:Oligonucleotides; D010455:Peptides; D011108:Polymers; D020033:Protein Isoforms; D011506:Proteins
keywords: Biomolecule analysis; Capillary columns; Enzymatic reactors; Review; Spatial chromatography; Stationary phases; Two-dimensional liquid chromatography