Citation: LI Jiabin, ZHOU Lianqi, YAN Hui, LI Nannan, HAO Feiran, TIAN Fang, ZHANG Yangjun. Metal-tag labeling coupled with high performance liquid chromatography-selected ion monitoring mass spectrometry for absolute quantitation of proteins[J]. Chinese Journal of Chromatography, ;2014, 32(4): 361-368. doi: 10.3724/SP.J.1123.2013.11042 shu

Metal-tag labeling coupled with high performance liquid chromatography-selected ion monitoring mass spectrometry for absolute quantitation of proteins

1.
State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, Beijing 102206, China

Corresponding author: ZHANG Yangjun,
Received Date: 22 November 2013
Available Online: 2 January 2014
Fund Project: 国家重大科学计划项目(2012CB910603，2010CB912704)；国家高技术研究发展规划(2012AA020202)；国家重大科学仪器设备开发专项项目(2011YQ030139，2011YQ06008408，2011YQ09000504，2012YQ12004407)；国家自然科学基金项目(20735005，31100591，21275159). (2012CB910603，2010CB912704)；国家高技术研究发展规划(2012AA020202)；国家重大科学仪器设备开发专项项目(2011YQ030139，2011YQ06008408，2011YQ09000504，2012YQ12004407)；国家自然科学基金项目(20735005，31100591，21275159)

A novel method has been established based on metal element chelated tags coupled with high performance liquid chromatography-selected ion monitoring mass spectrometry (HPLC-SIM/MS). The labeling efficiency and stability of metal element chelated tags, the chromatographic retention behavior and MS behavior of the labeled peptides, the linear range and accuracy of this method were examined. The results showed that the metal element chelated tag method has high labeling efficiency and high labeling stability, and the labeled peptides with different kinds of metal tags have consistent chromatographic retention behavior. The method of metal tags coupled with HPLC-SIM/MS has high sensitivity with the limit of quantification (LOQ) up to 1 fmol. The linear range for the method was between 1 fmol to 500 fmol with R²>0.99, which means the method has a good linearity. Moreover, this method had an average recovery of 117.01%. The method was used in the absolute quantitation of a protein enolase in Thermoanaerobacter tengcongensis (TTE) with a relative standard deviation of 5.74%, which means high precision. All the results showed that this method is accurate and reliable for the absolute quantitation of proteins. This gives us an alternative for the quantitative determination of proteins in relatively simple biological samples.
- metal labeling,
- high performance liquid chromatography (HPLC),
- selected ion monitoring mass spectrometry (SIM/MS),
- proteins,
- absolute quantitation
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