title: Capillary zone electrophoresis-multiple reaction monitoring from 100 pg of RAW 264.7 cell lysate digest.
abstract: Capillary zone electrophoresis-multiple/single reaction monitoring (CZE-MRM/SRM), which employed an electrokinetically driven sheath-flow electrospray interface, was used for the rapid and highly sensitive detection of protein analytes in complex tryptic digests. MRM channels were developed against a commercial exponential mixture of bovine proteins. Five proteins spanning four orders of magnitude concentration range were confidently detected from only 2.5 ng of the digest mixture; the mass detection limits (S/N = 3) of two detected proteins, alpha-casein and glutamate dehydrogenase were about 600 zmol and 30 amol, respectively. This technique was then applied to a RAW 264.7 cell lysate digest. Three proteins were confidently and reproducibly detected from 100 pg of this digest. The sample amount corresponds to the approximate protein content from a single cell, which suggests that CZE-MRM may be a useful analytical tool in chemical cytometry. In addition to providing highly sensitive detection of proteins in complex mixtures, this system is highly rapid; migration time of the protein digests was less than 10 min.
authors.affiliation: Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, IN 46556, USA.
authors.forename: Liangliang
authors.initials: L
authors.lastname: Sun
authors.forename: Yihan
authors.initials: Y
authors.lastname: Li
authors.forename: Matthew M
authors.initials: MM
authors.lastname: Champion
authors.forename: Guijie
authors.initials: G
authors.lastname: Zhu
authors.forename: Roza
authors.initials: R
authors.lastname: Wojcik
authors.forename: Norman J
authors.initials: NJ
authors.lastname: Dovichi
mesh_terms: D000818:Animals; D002417:Cattle; D016923:Cell Death; D002460:Cell Line; D019075:Electrophoresis, Capillary / Q000379:methods*; D051379:Mice; D011506:Proteins / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism; D059748:Proteolysis; D015203:Reproducibility of Results; D013997:Time Factors; D014357:Trypsin / Q000378:metabolism*
chemical_list: D011506:Proteins; D014357:Trypsin