Unknown

Dataset Information

0

Facile trypsin immobilization in polymeric membranes for rapid, efficient protein digestion.


ABSTRACT: Sequential adsorption of poly(styrene sulfonate) and trypsin in nylon membranes provides a simple, inexpensive method to create stable, microporous reactors for fast protein digestion. The high local trypsin concentration and short radial diffusion distances in membrane pores facilitate proteolysis in residence times of a few seconds, and the minimal pressure drop across the thin membranes allows their use in syringe filters. Membrane digestion and subsequent MS analysis of bovine serum albumin provide 84% sequence coverage, which is higher than the 71% coverage obtained with in-solution digestion for 16 h or the <50% sequence coverages of other methods that employ immobilized trypsin. Moreover, trypsin-modified membranes digest protein in the presence of 0.05 wt % sodium dodecyl sulfate (SDS), whereas in-solution digestion under similar conditions yields no peptide signals in mass spectra even after removal of SDS. These membrane reactors, which can be easily prepared in any laboratory, have a shelf life of several months and continuously digest protein for at least 33 h without significant loss of activity.

SUBMITTER: Xu F 

PROVIDER: S-EPMC3052767 | biostudies-literature | 2010 Dec

REPOSITORIES: biostudies-literature

altmetric image

Publications

Facile trypsin immobilization in polymeric membranes for rapid, efficient protein digestion.

Xu Fei F   Wang Wei-Han WH   Tan Yu-Jing YJ   Bruening Merlin L ML  

Analytical chemistry 20101118 24


Sequential adsorption of poly(styrene sulfonate) and trypsin in nylon membranes provides a simple, inexpensive method to create stable, microporous reactors for fast protein digestion. The high local trypsin concentration and short radial diffusion distances in membrane pores facilitate proteolysis in residence times of a few seconds, and the minimal pressure drop across the thin membranes allows their use in syringe filters. Membrane digestion and subsequent MS analysis of bovine serum albumin  ...[more]

Similar Datasets

| S-EPMC8804297 | biostudies-literature
| S-EPMC7671371 | biostudies-literature
| S-EPMC6961038 | biostudies-literature
| S-EPMC3253205 | biostudies-literature
| S-EPMC7377536 | biostudies-literature
| S-EPMC8743033 | biostudies-literature
| S-EPMC7406678 | biostudies-literature
| S-EPMC4563727 | biostudies-literature
| S-EPMC2950708 | biostudies-literature
| S-EPMC10059984 | biostudies-literature