PMID- 31800120
OWN - NLM
STAT- MEDLINE
DCOM- 20210723
LR  - 20210723
IS  - 1097-0231 (Electronic)
IS  - 0951-4198 (Print)
IS  - 0951-4198 (Linking)
VI  - 34 Suppl 4
IP  - Suppl 4
DP  - 2020 Sep
TI  - The measurement of KRAS G12 mutants using multiplexed selected reaction 
      monitoring and ion mobility mass spectrometry.
PG  - e8657
LID - 10.1002/rcm.8657 [doi]
LID - e8657
AB  - RATIONALE: There is a considerable clinical demand to determine key mutations in 
      genes involved with cancer which necessitates the deployment of highly specific 
      and robust analytical methods. Multiplex liquid chromatography with selected 
      reaction monitoring (LC/SRM) assays offer the ability to achieve quantitation 
      down to levels expected to be present in clinical samples. Ion mobility mass 
      spectrometry (IMS/MS) assays can provide increased peak capacity and hence 
      separation in an extremely short time frame, and in addition provide 
      physicochemical data regarding the collision cross-section of an analyte which 
      can be used in conjunction with the m/z value of an ion to increase detection 
      specificity. METHODS: For LC/SRM, unlabelled peptides and corresponding 
      stable-isotope-labelled standards were spiked into digested human plasma and 
      analysed using ultrahigh-performance liquid chromatography (UHPLC) coupled to a 
      triple quadrupole mass spectrometer to enable the generation of analyte-specific 
      calibration lines. Synthetic unlabelled peptides were infused into a Synapt G2 
      mass spectrometer for travelling wave ion mobility separation and (TW) CCS(N2) 
      values were derived from comparison with previously generated (TW) CCS(N2) 
      calibration values. RESULTS: Linear calibration lines (0.125 to 25 fmol/muL) were 
      established for each of the KRAS peptides. UHPLC separated the peptides and hence 
      enabled them to be split into different retention time functions/windows. This 
      separation enabled detection of three or four transitions for each light and 
      heavy peptide with at least 10 points per peak for accurate quantitation. All six 
      KRAS G12 peptides were separated using IMS/MS, enabling precise (TW) CCS(N2) 
      values to be determined. Although some of the G12 peptides chromatographically 
      co-eluted, all the peptides were distinguished by m/z, retention time and/or 
      drift time. CONCLUSIONS: This study advocates that LC/SRM and IMS/MS could both 
      be used to identify single amino acid substitutions in KRAS as an alternative to 
      commonly used methods such as circulating tumour DNA analysis.
CI  - (c) 2019 The Authors. Rapid Communications in Mass Spectrometry published by John 
      Wiley & Sons Ltd.
FAU - Norman, Rachel L
AU  - Norman RL
AD  - Leicester Cancer Research Centre, Leicester Royal Infirmary, University of 
      Leicester, Leicester, LE1 5WW, UK.
FAU - Singh, Rajinder
AU  - Singh R
AD  - Leicester Cancer Research Centre, Leicester Royal Infirmary, University of 
      Leicester, Leicester, LE1 5WW, UK.
FAU - Langridge, James I
AU  - Langridge JI
AD  - Waters Corporation, Wilmslow SK9 4AX, UK.
FAU - Ng, Leong L
AU  - Ng LL
AD  - Department of Cardiovascular Sciences, University of Leicester and National 
      Institute for Health Research Leicester Biomedical Research Centre, Glenfield 
      Hospital, Leicester, LE1 7RH, UK.
FAU - Jones, Donald J L
AU  - Jones DJL
AUID- ORCID: 0000-0001-6583-870X
AD  - Leicester Cancer Research Centre, Leicester Royal Infirmary, University of 
      Leicester, Leicester, LE1 5WW, UK.
AD  - Department of Cardiovascular Sciences, University of Leicester and National 
      Institute for Health Research Leicester Biomedical Research Centre, Glenfield 
      Hospital, Leicester, LE1 7RH, UK.
LA  - eng
PT  - Journal Article
DEP - 20200214
PL  - England
TA  - Rapid Commun Mass Spectrom
JT  - Rapid communications in mass spectrometry : RCM
JID - 8802365
RN  - 0 (KRAS protein, human)
RN  - 0 (Peptide Fragments)
RN  - EC 3.6.5.2 (Proto-Oncogene Proteins p21(ras))
SB  - IM
MH  - Amino Acid Substitution
MH  - Chromatography, High Pressure Liquid
MH  - Humans
MH  - Ion Mobility Spectrometry
MH  - Mass Spectrometry/*methods
MH  - *Mutation
MH  - Peptide Fragments/analysis/chemistry/metabolism
MH  - *Proto-Oncogene Proteins p21(ras)/blood/chemistry
PMC - PMC7539944
EDAT- 2019/12/05 06:00
MHDA- 2021/07/24 06:00
PMCR- 2020/10/07
CRDT- 2019/12/05 06:00
PHST- 2019/07/22 00:00 [received]
PHST- 2019/10/02 00:00 [revised]
PHST- 2019/11/09 00:00 [accepted]
PHST- 2019/12/05 06:00 [pubmed]
PHST- 2021/07/24 06:00 [medline]
PHST- 2019/12/05 06:00 [entrez]
PHST- 2020/10/07 00:00 [pmc-release]
AID - RCM8657 [pii]
AID - 10.1002/rcm.8657 [doi]
PST - ppublish
SO  - Rapid Commun Mass Spectrom. 2020 Sep;34 Suppl 4(Suppl 4):e8657. doi: 
      10.1002/rcm.8657. Epub 2020 Feb 14.