Biopolymer Chromatography on Inorganic Monoliths

In the field of biopolymer chromatography, monolithic columns, based on copolymerization of silane precursors, have predominately been investigated regarding separation of peptides and complex protein digests, whereas their application to analysis of high-molecules-weight analytes like proteins or dsDNA fragments is scarcely reported. 

In addition to classical reverse phase separation of peptides on octadecyl derivatized silica monoliths, sugars and peptides as well as proteins and nucleosides have been analyzed on a 20-cm-long silica-based poly(acrylic acid) column (ID. 200 pm), employing HILIC and weak cation-exchange chromatography, respectively [194]. Furthermore, HILIC fractionation of polysaccharides delivered remarkable and promising results [84,194]. 

On-line hydrolysis of proteins catalyzed by trypsin or pepsin immobilized on monolithic silica beds was described by Kato et al. [86,195], whereas pepsin was encapsnlated into the silica-gel matrix (75 pm capillary column), without loss in enzymatic activity [195]. 

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