title: A database of reaction monitoring mass spectrometry assays for elucidating therapeutic response in cancer.
abstract: OBJECTIVE
  The Quantitative Assay Database (QuAD), http://proteome.moffitt.org/QUAD/, facilitates widespread implementation of quantitative mass spectrometry in cancer biology and clinical research through sharing of methods and reagents for monitoring protein expression and modification.
  
  
  METHODS
  Liquid chromatography coupled to multiple reaction monitoring (LC-MRM) mass spectrometry assays are developed using SDS-PAGE fractionated lysates from cancer cell lines. Pathway maps created using GeneGO Metacore provide the biological relationships between proteins and illustrate concepts for multiplexed analysis; each protein can be selected to examine assay development at the protein and peptide levels.
  
  
  RESULTS
  The coupling of SDS-PAGE and multiple reaction monitoring mass spectrometry screening has been used to detect 876 peptides from 218 cancer-related proteins in model systems including colon, lung, melanoma, leukemias, and myeloma, which has led to the development of 95 quantitative assays including stable-isotope-labeled peptide standards. Methods are published online and peptide standards are made available to the research community. Protein expression measurements for heat shock proteins, including a comparison with ELISA and monitoring response to the HSP90 inhibitor, 17-(dimethylaminoethylamino)-17-demethoxygeldanamycin (17-DMAG), are used to illustrate the components of the QuAD and its potential utility.
  
  
  CONCLUSIONS
  This resource enables quantitative assessment of protein components of signaling pathways and biological processes and holds promise for systematic investigation of treatment responses in cancer.
authors.affiliation: Molecular Oncology and Proteomics, H. Lee Moffitt Cancer Center, Tampa, FL 33612, USA.
authors.forename: Elizabeth R
authors.initials: ER
authors.lastname: Remily-Wood
authors.forename: Richard Z
authors.initials: RZ
authors.lastname: Liu
authors.forename: Yun
authors.initials: Y
authors.lastname: Xiang
authors.forename: Yi
authors.initials: Y
authors.lastname: Chen
authors.forename: C Eric
authors.initials: CE
authors.lastname: Thomas
authors.forename: Neal
authors.initials: N
authors.lastname: Rajyaguru
authors.forename: Laura M
authors.initials: LM
authors.lastname: Kaufman
authors.forename: Joana E
authors.initials: JE
authors.lastname: Ochoa
authors.forename: Lori
authors.initials: L
authors.lastname: Hazlehurst
authors.forename: Javier
authors.initials: J
authors.lastname: Pinilla-Ibarz
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Lancet
authors.forename: Guolin
authors.initials: G
authors.lastname: Zhang
authors.forename: Eric
authors.initials: E
authors.lastname: Haura
authors.forename: David
authors.initials: D
authors.lastname: Shibata
authors.forename: Timothy
authors.initials: T
authors.lastname: Yeatman
authors.forename: Keiran S M
authors.initials: KS
authors.lastname: Smalley
authors.forename: William S
authors.initials: WS
authors.lastname: Dalton
authors.forename: Emina
authors.initials: E
authors.lastname: Huang
authors.forename: Ed
authors.initials: E
authors.lastname: Scott
authors.forename: Gregory C
authors.initials: GC
authors.lastname: Bloom
authors.forename: Steven A
authors.initials: SA
authors.lastname: Eschrich
authors.forename: John M
authors.initials: JM
authors.lastname: Koomen
mesh_terms: D000970:Antineoplastic Agents / Q000008:administration & dosage* / Q000627:therapeutic use; D016227:Benzoquinones / Q000494:pharmacology; D001681:Biological Assay; D002853:Chromatography, Liquid / Q000379:methods; D016208:Databases, Factual / Q000592:standards / Q000600:supply & distribution; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D018841:HSP90 Heat-Shock Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007553:Isotope Labeling; D047029:Lactams, Macrocyclic / Q000494:pharmacology; D013058:Mass Spectrometry / Q000379:methods; D009369:Neoplasms / Q000175:diagnosis* / Q000188:drug therapy / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D010455:Peptides / Q000032:analysis* / Q000737:chemistry; D011379:Prognosis; D040901:Proteomics / Q000295:instrumentation / Q000379:methods; D012015:Reference Standards; D015398:Signal Transduction / Q000235:genetics
chemical_list: D000970:Antineoplastic Agents; D016227:Benzoquinones; D018841:HSP90 Heat-Shock Proteins; D047029:Lactams, Macrocyclic; D010455:Peptides; C448659:17-(dimethylaminoethylamino)-17-demethoxygeldanamycin