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[Identification rhFKBP12 by ESI-quadrupole-oa-TOF tandem mass spectrometry]
Hong-xia Wang1, Xue-min Zhang, Song-cheng Yang
1National Center of Biomedical Analysis, Academy of Military Medical Sciences, Beijing 100850, China.
Aim:
To identify recombinant protein rhFKBP12 by new technique ESI-quadrupole-oa-TOF tandem mass spectrometry.
Methods:
The molecular weight of rhFKBP12 was measured by ESI-MS. Digest rhFKBP12 by trypsin at 37 degrees C over night. The tryptic digested peptides were measured and then two doublely charged peptides were selected to measure their amino acid sequence by ESI-MS/MS. Search database with the measured amino acid sequence to identify rhFKBP12.
Results:
The calculated molecular weight of rhFKBP12 was 11,819.54 and the measured value was 11,820.38. The measurement percent error was only 0.007%. The sequence measured by ESI-MS/MS was QVETMS and EEGVAQMSV and then the database search results with them were both hFKBP12.
Conclusion:
The study proves that the primary structure of rhFKBP12 is correct and there is no amino acid deletion, mutation and modification in its expression, refolding and purification. It also shows that ESI-MS/MS is a good method to identify protein with advantage of sensitivity, high speed and accuracy.