Biocompatible polymeric monoliths for protein and peptide separations

Abstract

The concept of biocompatibility with reference to chromatographic stationary phases for separation of biomolecules (including proteins and peptides) is introduced. Biocompatible is a characteristic that indicates resistance to nonspecific adsorption of biomolecules and preservation of their structures and biochemical functions. Two types of biocompatible polymeric monoliths [i. e., polyacrylamide- and poly(meth)acrylate-based monoliths] used for protein and peptide separations are reviewed in detail, with emphasis on size exclusion, ion exchange, and hydrophobic interaction chromatographic modes. Biocompatible monoliths for enzyme reactors are also included. The two main synthetic approaches to produce biocompatible monoliths are summarized, i. e., surface modification of a monolith that is not inherently biocompatible and direct copolymerization of hydrophilic monomers to form a biocompatible monolith directly. Integration of polyethylene glycol into the poly(meth)acrylate monolith network is becoming popular for reduction of non-specific protein interactions.

An Open Tubular CEC Column of Excellent Separation Efficiency for Proteomic Analysis

“…The typical micro-or nano-LC column packed with ca 3 µm C18 particles shows generally good separation performance, but the practical column length has been less than 20 cm owing to high column back pressure, thus its overall column separation efficiency has been limited. Monolithic columns may be adopted instead, 17,18 but the overall column separation efficiency has been limited, too.…”

Collection of Peptides Released from Single Neurons with Particle-Embedded Monolithic Capillaries Followed by Detection with Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry

“…20–22 One advantage of these materials is the ease with which the chemical properties of the monolith can be optimized for specific analyte classes, either by selecting different monomers/cross-linkers or further modifying the monolith with the intended functional group after polymerization. 23–26, This results in the availability of multiple surface chemistries and improved performance in extraction or separation of specific analytes.…”

Sequence-specific DNA solid-phase extraction in an on-chip monolith: Towards detection of antibiotic resistance genes

“…In initial studies, ethylene dimethacrylate was used as a crosslinker; however, this monolith formulation suffered from nonspecific adsorption of fluorescent molecules when bacterial lysate samples were introduced. Monoliths prepared with PEGDA as an alternate, more hydrophilic crosslinker [26] showed much less nonspecific adsorption and were therefore used for the reported experiments. The ratio of the porogens, 1-propanol and 1,4-butanediol, was studied regarding monolith morphology (evaluated by SEM) and flow-through properties.…”