PMID- 17221927
OWN - NLM
STAT- MEDLINE
DCOM- 20070509
LR  - 20181201
IS  - 1076-5174 (Print)
IS  - 1076-5174 (Linking)
VI  - 42
IP  - 2
DP  - 2007 Feb
TI  - Automated mass correction and data interpretation for protein open-access liquid 
      chromatography-mass spectrometry.
PG  - 139-49
AB  - Characterization of recombinant protein purification fractions and final products 
      by liquid chromatography-mass spectrometry (LC/MS) are requested more frequently 
      each year. A protein open-access (OA) LC/MS system was developed in our 
      laboratory to meet this demand. This paper compares the system that we originally 
      implemented in our facilities in 2003 to the one now in use, and discusses, in 
      more detail, recent enhancements that have improved its robustness, reliability, 
      and data reporting capabilities. The system utilizes instruments equipped with 
      reversed-phase chromatography and an orthogonal accelerated time-of-flight mass 
      spectrometer fitted with an electrospray source. Sample analysis requests are 
      accomplished using a simple form on a web-enabled laboratory information 
      management system (LIMS). This distributed form is accessible from any 
      intranet-connected company desktop computer. Automated data acquisition and 
      processing are performed using a combination of in-house (OA-Self Service, 
      OA-Monitor, and OA-Analysis Engine) and vendor-supplied programs (AutoLynx, and 
      OpenLynx) located on acquisition computers and off-line processing workstations. 
      Analysis results are then reported via the same web-based LIMS. Also presented 
      are solutions to problems not addressed on commercially available, small-molecule 
      OA-LC/MS systems. These include automated transforming of mass-to-charge (m/z) 
      spectra to mass spectra and automated data interpretation that considers minor 
      variants to the protein sequence-such as common post-translational modifications 
      (PTMs). Currently, our protein OA-LC/MS platform runs on five LC/MS instruments 
      located in three separate GlaxoSmithKline R&D sites in the US and UK. To date, 
      more than 8000 protein OA-LC/MS samples have been analyzed. With these user 
      friendly and highly automated OA systems in place, mass spectrometry plays a key 
      role in assessing the quality of recombinant proteins, either produced at our 
      facilities or bought from external sources, without dedicating extensive amounts 
      of analyst resource.
CI  - Copyright 2007 John Wiley & Sons, Ltd.
FAU - Wagner, Craig D
AU  - Wagner CD
AD  - Molecular Discovery Research, GlaxoSmithKline, Research Triangle Park, NC 27709, 
      USA.
FAU - Hall, John T
AU  - Hall JT
FAU - White, Wendy L
AU  - White WL
FAU - Miller, Luke A D
AU  - Miller LA
FAU - Williams, Jon D
AU  - Williams JD
LA  - eng
PT  - Comparative Study
PT  - Journal Article
PL  - England
TA  - J Mass Spectrom
JT  - Journal of mass spectrometry : JMS
JID - 9504818
RN  - 0 (Caseins)
RN  - 0 (Recombinant Proteins)
RN  - EC 4.2.1.1 (Carbonic Anhydrases)
SB  - IM
MH  - Animals
MH  - Carbonic Anhydrases/*chemistry
MH  - Caseins/*chemistry
MH  - Cattle
MH  - Chromatography, High Pressure Liquid
MH  - *Electronic Data Processing
MH  - *Information Management
MH  - Peptide Mapping
MH  - Recombinant Proteins/chemistry
MH  - Reproducibility of Results
MH  - Spectrometry, Mass, Electrospray Ionization/instrumentation/*methods
EDAT- 2007/01/16 09:00
MHDA- 2007/05/10 09:00
CRDT- 2007/01/16 09:00
PHST- 2007/01/16 09:00 [pubmed]
PHST- 2007/05/10 09:00 [medline]
PHST- 2007/01/16 09:00 [entrez]
AID - 10.1002/jms.1174 [doi]
PST - ppublish
SO  - J Mass Spectrom. 2007 Feb;42(2):139-49. doi: 10.1002/jms.1174.