Human Serum Proteins Preseparated by Electrophoresis or Chromatography Followed by Tandem Mass Spectrometry

• Published in: Journal of proteome research Vol. 3; no. 3; pp. 364 - 382
• Main Authors: Marshall, John, Jankowski, Andy, Furesz, Shirley, Kireeva, Inga, Barker, Lisa, Dombrovsky, Mila, Zhu, Weimin, Jacks, Kellie, Ingratta, Leslee, Bruin, Jenny, Kristensen, Erika, Zhang, Rulin, Stanton, Eric, Takahashi, Miyoko, Jackowski, George
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 01.05.2004
• Subjects: Blood Proteins - chemistry; Chromatography - methods; Electrophoresis, Polyacrylamide Gel - methods; Humans; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods
• ISSN: 1535-3893; 1535-3907
• DOI: 10.1021/pr034039p

Electrophoretic and chromatographic sample preparations were compared and together detected the presence of some 600 types of protein products in human serum. Proteins from crude serum preseparated by ionic electrophoresis, chromatography, or a combination of both were analyzed. Proteins were digested with trypsin or chymotrypsin. Naturally occurring peptides were also collected by reversed-phase chromatography. The resulting peptides were identified by tandem mass spectrometry. The peptides were either desorbed by a laser from a metal chip into a quadrupole-time-of-flight mass spectrometer or ionized as an electro-spray from reversed-phase chromatography via a metal needle under voltage into an ion-trap mass spectrometer. All of the commonly known proteins associated with serum were detected, and the two mass spectrometers agreed on the identity of abundant serum proteins. Preseparation of serum proteins prior to digestion markedly enhanced the capacity to detect un-common proteins from blood. Electrophoretic- and chromatography-based experiments were found to be complementary. Many novel cellular proteins not previously associated with serum were recorded. Keywords: human • serum • protein • proteome • electrophoresis • chromatography • tandem • mass spectrometer