A preliminary study on screening for Porphyromonas gingivalis outer membrane protein antigen with two-dimensional liquid phase fractionation and matrix-assisted laser desorption ionization time-of-flight/time-of-flight mass spectrometry
10.3760/cma.j.issn.1002-0098.2010.12.012
- VernacularTitle:联合应用二维液相色谱及串联质谱技术筛选牙龈卟啉单胞菌外膜蛋白抗原的初步研究
- Author:
Ang LI
1
;
Wei-Hang SI
;
Si-Cen WANG
;
Jian-Feng SHI
;
Guo-Zhou RAO
;
Jian-Zhong GOU
Author Information
1. 西安交通大学医学院附属口腔医院
- Keywords:
Porphyromonas gingivalis;
Bacterial outer membrane proteins;
Chromatography liquid;
Matrix-assisted laser desorption ionization time-of-flight/time-of-flight mass spectrometry
- From:
Chinese Journal of Stomatology
2010;45(12):749-753
- CountryChina
- Language:Chinese
-
Abstract:
Objective To screen a variety of Porphyromonas gingivalis (Pg) common outer membrane proteins with two-dimensional liquid phase fractionation (PF2D) and matrix-assisted laser desorption ionization time-of-flight/time-of-flight mass spectrometry ( MALDI-TOF/TOF-MS ) and provide candidate target antigen for the design of vaccines with cross protection against a variety of Pg. Methods The outer membrane proteins of Pg301, PgATCC33277 and PgW83 were extracted through ultracentrifugation, and then they were separated by ProteomeLab PF2D protein fractionation system. After separation, the outer membrane proteins were obtained through comparison, and the primary structure of the proteins was identified by MALDI-TOF/TOF-MS and database. Results Ninety-nine protein samples out of 3 strains of Pg were obtained after the high performance chromato focusing (HPCF) separation process. B7 fractions of 3 strains of Pg were separated by the reversed-phase high performance liquid chromatography (RP-HPCL) separation process. After comparison of peak and retention time of chromatogram, the 8 common protein peaks of 3 strains of Pg were confirmed. The protein samples were identified by MALDI-TOF/TOF-MS, and one of them was known protein arg-gingipainA. Conclusions PF2D protein fractionation system is of good reproducibility and high resolution. A combination of PF2D and MALDI-TOF/TOF-MS can be used to identify the common outer memberane proteins of Pg.
