title: Industrial-scale proteomics: from liters of plasma to chemically synthesized proteins.
abstract: Human blood plasma is a useful source of proteins associated with both health and disease. Analysis of human blood plasma is a challenge due to the large number of peptides and proteins present and the very wide range of concentrations. In order to identify as many proteins as possible for subsequent comparative studies, we developed an industrial-scale (2.5 liter) approach involving sample pooling for the analysis of smaller proteins (M(r) generally < ca. 40 000 and some fragments of very large proteins). Plasma from healthy males was depleted of abundant proteins (albumin and IgG), then smaller proteins and polypeptides were separated into 12 960 fractions by chromatographic techniques. Analysis of proteins and polypeptides was performed by mass spectrometry prior to and after enzymatic digestion. Thousands of peptide identifications were made, permitting the identification of 502 different proteins and polypeptides from a single pool, 405 of which are listed here. The numbers refer to chromatographically separable polypeptide entities present prior to digestion. Combining results from studies with other plasma pools we have identified over 700 different proteins and polypeptides in plasma. Relatively low abundance proteins such as leptin and ghrelin and peptides such as bradykinin, all invisible to two-dimensional gel technology, were clearly identified. Proteins of interest were synthesized by chemical methods for bioassays. We believe that this is the first time that the small proteins in human blood plasma have been separated and analyzed so extensively.
authors.affiliation: GeneProt, 2 rue Pré-de-la-Fontaine, CP125, CH-1217, Meyrin 2, Switzerland. keith.rose@geneprot.com
authors.forename: Keith
authors.initials: K
authors.lastname: Rose
authors.forename: Lydie
authors.initials: L
authors.lastname: Bougueleret
authors.forename: Thierry
authors.initials: T
authors.lastname: Baussant
authors.forename: Günter
authors.initials: G
authors.lastname: Böhm
authors.forename: Paolo
authors.initials: P
authors.lastname: Botti
authors.forename: Jacques
authors.initials: J
authors.lastname: Colinge
authors.forename: Isabelle
authors.initials: I
authors.lastname: Cusin
authors.forename: Hubert
authors.initials: H
authors.lastname: Gaertner
authors.forename: Anne
authors.initials: A
authors.lastname: Gleizes
authors.forename: Manfred
authors.initials: M
authors.lastname: Heller
authors.forename: Silvia
authors.initials: S
authors.lastname: Jimenez
authors.forename: Andrew
authors.initials: A
authors.lastname: Johnson
authors.forename: Martin
authors.initials: M
authors.lastname: Kussmann
authors.forename: Laure
authors.initials: L
authors.lastname: Menin
authors.forename: Christoph
authors.initials: C
authors.lastname: Menzel
authors.forename: Frederic
authors.initials: F
authors.lastname: Ranno
authors.forename: Patricia
authors.initials: P
authors.lastname: Rodriguez-Tomé
authors.forename: John
authors.initials: J
authors.lastname: Rogers
authors.forename: Cedric
authors.initials: C
authors.lastname: Saudrais
authors.forename: Matteo
authors.initials: M
authors.lastname: Villain
authors.forename: Diana
authors.initials: D
authors.lastname: Wetmore
authors.forename: Amos
authors.initials: A
authors.lastname: Bairoch
authors.forename: Denis
authors.initials: D
authors.lastname: Hochstrasser
mesh_terms: D000595:Amino Acid Sequence; D001774:Blood Chemical Analysis / Q000379:methods*; D001798:Blood Proteins / Q000378:metabolism*; D002845:Chromatography; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D019295:Computational Biology; D019992:Databases as Topic; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry; D010949:Plasma / Q000378:metabolism*; D020543:Proteome; D040901:Proteomics / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013347:Subcellular Fractions; D013997:Time Factors; D014357:Trypsin / Q000494:pharmacology
chemical_list: D001798:Blood Proteins; D007074:Immunoglobulin G; D010455:Peptides; D020543:Proteome; D014357:Trypsin