title: Rapid development of sensitive, high-throughput, quantitative and highly selective mass spectrometric targeted immunoassays for clinically important proteins in human plasma and serum.
abstract: OBJECTIVE
  The aim of this study was to develop high-throughput, quantitative and highly selective mass spectrometric, targeted immunoassays for clinically important proteins in human plasma or serum.
  
  
  METHODS
  The described method coupled mass spectrometric immunoassay (MSIA), a previously developed technique for immunoenrichment on a monolithic microcolumn activated with an anti-protein antibody and fixed in a pipette tip, to selected reaction monitoring (SRM) detection and accurate quantification of targeted peptides, including clinically relevant sequence or truncated variants.
  
  
  RESULTS
  In this report, we demonstrate the rapid development of MSIA-SRM assays for sixteen different target proteins spanning seven different clinically important areas (including neurological, Alzheimer's, cardiovascular, endocrine function, cancer and other diseases) and ranging in concentration from pg/mL to mg/mL. The reported MSIA-SRM assays demonstrated high sensitivity (within published clinical ranges), precision, robustness and high-throughput as well as specific detection of clinically relevant isoforms for many of the target proteins. Most of the assays were tested with bona-fide clinical samples. In addition, positive correlations, (R2 0.67-0.87, depending on the target peptide), were demonstrated for MSIA-SRM assay data with clinical analyzer measurements of parathyroid hormone (PTH) and insulin growth factor 1 (IGF1) in clinical sample cohorts.
  
  
  CONCLUSIONS
  We have presented a practical and scalable method for rapid development and deployment of MS-based SRM assays for clinically relevant proteins and measured levels of the target analytes in bona fide clinical samples. The method permits the specific quantification of individual protein isoforms and addresses the difficult problem of protein heterogeneity in clinical proteomics applications.
authors: ThermoFisher Scientific BRIMS, 790 Memorial Dr, Cambridge, MA, USA.
authors: Bryan
authors: B
authors: Krastins
authors: Amol
authors: A
authors: Prakash
authors: David A
authors: DA
authors: Sarracino
authors: Dobrin
authors: D
authors: Nedelkov
authors: Eric E
authors: EE
authors: Niederkofler
authors: Urban A
authors: UA
authors: Kiernan
authors: Randall
authors: R
authors: Nelson
authors: Maryann S
authors: MS
authors: Vogelsang
authors: Gouri
authors: G
authors: Vadali
authors: Alejandra
authors: A
authors: Garces
authors: Jennifer N
authors: JN
authors: Sutton
authors: Scott
authors: S
authors: Peterman
authors: Gregory
authors: G
authors: Byram
authors: Bruno
authors: B
authors: Darbouret
authors: Joëlle R
authors: JR
authors: Pérusse
authors: Nabil G
authors: NG
authors: Seidah
authors: Benoit
authors: B
authors: Coulombe
authors: Johan
authors: J
authors: Gobom
authors: Erik
authors: E
authors: Portelius
authors: Josef
authors: J
authors: Pannee
authors: Kaj
authors: K
authors: Blennow
authors: Vathany
authors: V
authors: Kulasingam
authors: Lewis
authors: L
authors: Couchman
authors: Caje
authors: C
authors: Moniz
authors: Mary F
authors: MF
authors: Lopez
mesh_terms: D000544:Alzheimer Disease / Q000097:blood; D001798:Blood Proteins / Q000302:isolation & purification*; D002318:Cardiovascular Diseases / Q000097:blood; D006130:Growth Disorders / Q000097:blood; D057166:High-Throughput Screening Assays; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D013058:Mass Spectrometry / Q000379:methods*; D009369:Neoplasms / Q000097:blood; D051437:Renal Insufficiency / Q000097:blood
chemical_list: D001798:Blood Proteins