Measurement of the amino acid sequence for the fusion protein FP3 with LC-MS/MS.
- Author:
Xiang LI
1
;
Xiang-Dong GAO
;
Lei TAO
;
De-Ning PEI
;
Ying GUO
;
Chun-Ming RAO
;
Jun-Zhi WANG
Author Information
1. Department of Recombinant Product, National Institutes for Food and Drug Control, Key Laboratory of the Ministry of Health for Research on Quality and Standardization of Biotech Products, Beijing 100050, China.
- Publication Type:Journal Article
- MeSH:
Amino Acid Sequence;
Chromatography, High Pressure Liquid;
Molecular Sequence Data;
Peptide Mapping;
Recombinant Fusion Proteins;
chemistry;
Spectrometry, Mass, Electrospray Ionization;
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization;
Tandem Mass Spectrometry;
Vascular Endothelial Growth Factor A;
antagonists & inhibitors;
chemistry
- From:
Acta Pharmaceutica Sinica
2012;47(2):216-222
- CountryChina
- Language:Chinese
-
Abstract:
The amino acid sequence of the fusion protein FP3 was measured by two types of LC-MS/MS and its primary structure was confirmed. After reduction and alkylation, the protein was digested with trypsin and glycosyl groups in glycopeptide were removed by PNGase F. The mixed peptides were separated by LC, then Q-TOF and Ion trap tandem mass spectrometry were used to measure b, y fragment ions of each peptide to analyze the amino acid sequence of fusion protein FP3. Seventy-six percent of full amino acid sequence of the fusion protein FP3 was measured by LC-ESI-Q-TOF with the remaining 24% completed by LC-ESI-Trap. As LC-MS and tandem mass spectrometry are rapid, sensitive, accurate to measure the protein amino acid sequence, they are important approach to structure analysis and identification of recombinant protein.
