Analysis of allergen protein in Macrobrachium by ultra-high performanceliquid chromatography coupled with quadrupole orbitrap mass spectrometry
10.19485/j.cnki.issn2096-5087.2020.10.009
- VernacularTitle:超高效液相色谱-四极杆静电场轨道离子阱质谱分析沼虾过敏原蛋白
- Author:
PAN Xiaodong
1
;
HUANG Baifen
;
CAI Zengxuan
;
WANG Mengli
;
HAN Jianlong
;
CHEN Qing
Author Information
1. Zhejiang Provincial Center for Disease Control and Prevention
- Publication Type:Journal Article
- Keywords:
ultra-high performance liquid chromatography quadrupole orbitrap mass spectrometry allergen protein shrimp characteristic peptide
- From:
Journal of Preventive Medicine
2020;32(10):1010-1012
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To establish the ultra-high performance liquid chromatography coupled with quadrupole orbitrap mass spectrometry ( UPLC-Q-Orbitrap-MS ) for the analysis of allergen protein in Macrobrachium.
Methods:Based on the strategy of bottom-up protein analysis, the proteins in Macrobrachium samples were extracted by Tris-HCl, digested by trypsin at 40 ℃ for 6 hours, separated by chromatography, and analyzed by Q-Orbitrap-MS spectrometry ( Full MS/dd-MS2, TopN=10 ) . Allergen proteins were identified with UniProt protein database and Proteome Discoverer software.
Results: Four kinds of allergen proteins were obtained, which were tropomyosin, arginine kinase, sarcoplasmic calcium binding protein and hemocyanin. The coverage rates of peptides in proteins were 53%, 36%, 12% and 12%, respectively. Post translation modifications were methylation of aspartic acid (D), deacylylation of aspartic acid ( N ) , glutamyl ammonia ( Q ) and oxidation of methionine ( M ) .
Conclusions:The UPLC-Q-Orbitrap-MS can identified abundant peptide and fragment information with high sensitivity and resolution, which provides a technical support for the analysis of shrimp allergens.
- Full text:超高效液相色谱-四极杆静电场轨道离子阱质谱分析沼虾过敏原蛋白.pdf
