title: Trypsin-based monolithic bioreactor coupled on-line with LC/MS/MS system for protein digestion and variant identification in standard solutions and serum samples.
abstract: The applicability of a trypsin-based monolithic bioreactor coupled on-line with LC/MS/MS for rapid proteolytic digestion and protein identification is here described. Dilute samples are passed through the bioreactor for generation of proteolytic fragments in less than 10 min. After digestion and peptide separation, electrospray ionization tandem mass spectrometry is used to generate a peptide map and to identify proteolytic peptides by correlating their fragmentation spectra with amino acid sequences from a protein database. By digesting picomoles of proteins sufficient data from ESI and MS/MS were obtained to unambiguously identify proteins alone and in serum samples. This approach was also extended to locate mutation sites in beta-lactoglobulin A and B variants.
authors.affiliation: Department of Pharmaceutical Chemistry, University of Pavia, Via Taramelli 12, I-27100 Pavia, Italy. enrica.calleri@unipv.it
authors.forename: Enrica
authors.initials: E
authors.lastname: Calleri
authors.forename: Caterina
authors.initials: C
authors.lastname: Temporini
authors.forename: Eleonora
authors.initials: E
authors.lastname: Perani
authors.forename: Antonella
authors.initials: A
authors.lastname: De Palma
authors.forename: Dieter
authors.initials: D
authors.lastname: Lubda
authors.forename: Giorgio
authors.initials: G
authors.lastname: Mellerio
authors.forename: Alberto
authors.initials: A
authors.lastname: Sala
authors.forename: Monica
authors.initials: M
authors.lastname: Galliano
authors.forename: Gabriele
authors.initials: G
authors.lastname: Caccialanza
authors.forename: Gabriella
authors.initials: G
authors.lastname: Massolini
mesh_terms: D000595:Amino Acid Sequence; D019149:Bioreactors; D002853:Chromatography, Liquid / Q000379:methods*; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D011506:Proteins / Q000737:chemistry*; D012015:Reference Standards; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D014357:Trypsin / Q000378:metabolism*
chemical_list: D011506:Proteins; D014357:Trypsin