Trypsin coatings on electrospun and alcohol-dispersed polymer nanofibers for a trypsin digestion column.

ABSTRACT: The construction of a trypsin column for rapid and efficient protein digestion in proteomics is described. Electrospun and alcohol-dispersed polymer nanofibers were used for the fabrication of highly stable trypsin coatings, which were prepared by a two-step process of covalent attachment and enzyme cross-linking. In a comparative study with the trypsin coatings on as-spun and nondispersed nanofibers, it has been observed that a simple step of alcohol dispersion improved not only the enzyme loading but also the performance of protein digestion. In-column digestion of enolase was successfully performed in less than 20 min. By applying the alcohol dispersion of polymer nanofibers, the bypass of samples was reduced by filling up the column with well-dispersed nanofibers, and subsequently, int

SUBMITTER: Jun SH 

PROVIDER: S-EPMC2950708 | biostudies-literature | 2010 Sep

REPOSITORIES: biostudies-literature