Identification of Bacterial Surface Antigens by Screening Peptide Phage Libraries Using Whole Bacteria Cell-Purified Antisera.
Bacterial surface proteins can be good vaccine candidates. In the present study, we used polyclonal antibodies purified with intactErysipelothrix rhusiopthiaeto screen phage-displayed random dodecapeptide and loop-constrained heptapeptide libraries, which led to the identification of mimotopes. Homology search of the mimotope sequences againstE. rhusiopthiae-encoded ORF sequences revealed 14 new antigens that may localize on the surface ofE. rhusiopthiae. When these putative surface proteins were used to immunize mice, 9/11 antigens induced protective immunity. Thus, we have demonstrated that a combination of using the whole bacterial cells to purify antibodies and using the phage-displayed peptide libraries to determine the antigen specificities of the antibodies can lead to the discovery of novel bacterial surface antigens. This can be a general approach for identifying surface antigens for other bacterial species.
College of Veterinary Medicine, Preventive Veterinary Medicine, Hunan Agricultural University Changsha, China.
Yun-Fei
YF
Hu
College of Veterinary Medicine, Preventive Veterinary Medicine, Hunan Agricultural University Changsha, China.
Dun
D
Zhao
College of Veterinary Medicine, Preventive Veterinary Medicine, Hunan Agricultural University Changsha, China.
Xing-Long
XL
Yu
College of Veterinary Medicine, Preventive Veterinary Medicine, Hunan Agricultural University Changsha, China.
Yu-Li
YL
Hu
College of Veterinary Medicine, Preventive Veterinary Medicine, Hunan Agricultural University Changsha, China.
Run-Cheng
RC
Li
College of Veterinary Medicine, Preventive Veterinary Medicine, Hunan Agricultural University Changsha, China.
Meng
M
Ge
College of Veterinary Medicine, Preventive Veterinary Medicine, Hunan Agricultural University Changsha, China.
Tian-Qi
TQ
Xu
College of Veterinary Medicine, Preventive Veterinary Medicine, Hunan Agricultural University Changsha, China.
Xiao-Bo
XB
Liu
College of Veterinary Medicine, Preventive Veterinary Medicine, Hunan Agricultural University Changsha, China.
Hua-Yuan
HY
Liao
Erysipelothrix rhusiopthiae; homology analysis; mimotope; phage display; subcellular localization; surface protein