A subtractive fluorescence-activated cell-sorting strategy to identify mimotopes of HBV-preS protein from bacterially displayed peptide library.
A novel subtractive fluorescence-activated cell-sorting (FACS) strategy using a model system is described here to identify disease-specific (DS) epitopes from a bacterially displayed random peptide library. In this process, preimmune serum was used as "Driver " to block any common binding sites on the bacterial surface and the labeled anti-preS IgG polyclonal antibodies from immunized serum were used as "Tester" to enrich preS-specific mimotopes. Bacterial clones were identified out of this pool through an "antigen-independent" procedure only using both different sera samples. After four rounds of sub-FACS screening, 41 out of 50 bacterial clones were identified as reacting with the immunized serum but not reacting with the pre-immune one. Two motif sequences HQLD and DPAF were obtained from 13 clones. Immunization of mice with two representative bacterial clones elicited a strong specific response against native preS antigen in comparison with the control. This technique may provide a useful technology platform for high-throughput screening of disease-related epitope which is of importance to develop vaccine against some infectious diseases whose pathogen or immunodominant antigen is still unknown.
Xin
Zhong-Tao
ZT
Department of Biochemistry, Beijing Institute of Basic Medical Sciences, P.O. Box 130 (3), Beijing, 100850, People's Republic of China.
Liu
Chuan
C
Dong
Bo
B
Gao
Ya-Ping
YP
Shao
Ning-Sheng
NS
Liu
Wei
W
Zhang
Jie
J
Dong
Jie
J
Ling
Shi-Gan
SG
Xue
Yan-Ning
YN
D000818:Animals; D015153:Blotting, Western; D002469:Cell Separation / Q000379:methods*; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D006515:Hepatitis B virus / Q000276:immunology*; D051379:Mice; D019151:Peptide Library; D011498:Protein Precursors / Q000276:immunology*; D011817:Rabbits; D014759:Viral Envelope Proteins / Q000276:immunology*
D019151:Peptide Library; D011498:Protein Precursors; D014759:Viral Envelope Proteins; C101581:preS (120-145) peptide