title: High-throughput peptide mass mapping using a microdevice containing trypsin immobilized on a porous polymer monolith coupled to MALDI TOF and ESI TOF mass spectrometers.
abstract: An enzymatic microreactor with a volume of 470 nL has been prepared by immobilizing trypsin on a 10 cm long reactive porous polymer monolith located in a 100 microm i.d. fused silica capillary. This reactor affords suitable degrees of digestion of proteins even after very short residence times of less than 1 min. The performance is demonstrated with the digestion of eight proteins ranging in molecular mass from 2848 to 77 754. The digests were analyzed using mass spectrometry in two modes: off-line MALDI and in-line nanoelectrospray ionization. The large numbers of identified peptides enable a high degree of sequence coverage and positive identification of the proteins. The extent of sequence coverage decreases as the molecular mass of the digested protein increases.
authors.affiliation: E. O. Lawrence Berkeley National Laboratory, Materials Sciences Division, Berkeley, California 94720, USA.
authors.forename: Dominic S
authors.initials: DS
authors.lastname: Peterson
authors.forename: Thomas
authors.initials: T
authors.lastname: Rohr
authors.forename: Frantisek
authors.initials: F
authors.lastname: Svec
authors.forename: Jean M J
authors.initials: JM
authors.lastname: Fréchet
mesh_terms: D010449:Peptide Mapping / Q000379:methods*; D011506:Proteins / Q000032:analysis / Q000378:metabolism; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014357:Trypsin / Q000378:metabolism*
chemical_list: D011506:Proteins; D014357:Trypsin