title: A subtractive fluorescence-activated cell-sorting strategy to identify mimotopes of HBV-preS protein from bacterially displayed peptide library.
abstract: 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.
authors.affiliation: Department of Biochemistry, Beijing Institute of Basic Medical Sciences, P.O. Box 130 (3), Beijing, 100850, People's Republic of China.
authors.forename: Zhong-Tao
authors.initials: ZT
authors.lastname: Xin
authors.forename: Chuan
authors.initials: C
authors.lastname: Liu
authors.forename: Bo
authors.initials: B
authors.lastname: Dong
authors.forename: Ya-Ping
authors.initials: YP
authors.lastname: Gao
authors.forename: Ning-Sheng
authors.initials: NS
authors.lastname: Shao
authors.forename: Wei
authors.initials: W
authors.lastname: Liu
authors.forename: Jie
authors.initials: J
authors.lastname: Zhang
authors.forename: Jie
authors.initials: J
authors.lastname: Dong
authors.forename: Shi-Gan
authors.initials: SG
authors.lastname: Ling
authors.forename: Yan-Ning
authors.initials: YN
authors.lastname: Xue
mesh_terms: 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*
chemical_list: D019151:Peptide Library; D011498:Protein Precursors; D014759:Viral Envelope Proteins; C101581:preS (120-145) peptide