title: Predefined spacers between epitopes on a recombinant epitope-peptide impacted epitope-specific antibody response.
abstract: We have developed a widely applicable method to construct epitope-peptide gene for epitope-vaccine strategy recently. In this study, we wanted to know whether the predefined spacers between epitopes on a recombinant epitope-peptide impacted the production of epitope-specific antibodies. The neutralizing epitope ELDKWA on the C-domain of HIV-1 gp41 was defined by the monoclonal antibody (mAb) 2F5 with broad neutralizing activity. We constructed three recombinant ELDKWA-epitope-peptides with different spacers between epitopes. The recombinant epitope-peptide GST-K8, GST-S8 and GST-R8 were bearing eight copies of ELDKWA-epitope with amino acid spacer GS, GSGGGGS and RS, respectively. GST-K8 and GST-S8 could induce high titer of ELDKWA-epitope-specific antibodies, much better than GST-R8. Besides, both antibodies could recognize the recombinant soluble gp41 and the transfected CHO-WT cells that stably express HIV-1 envelope glycoprotein on the cell surfaces. These experimental results indicated that the spacer GSGGGGS and GS were feasible in constructing a recombinant epitope-vaccine.
authors.affiliation: Laboratory of Immunology, Protein Science Laboratory of MOE, Department of Biology, Institute for Biomedical Science, Tsinghua University, Beijing 100084, PR China.
authors.forename: Zuqiang
authors.initials: Z
authors.lastname: Liu
authors.forename: Zuguang
authors.initials: Z
authors.lastname: Wang
authors.forename: Ying-Hua
authors.initials: YH
authors.lastname: Chen
mesh_terms: D000818:Animals; D000906:Antibodies / Q000235:genetics / Q000276:immunology*; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000235:genetics / Q000276:immunology*; D015151:Immunoblotting; D051379:Mice; D010455:Peptides / Q000235:genetics / Q000276:immunology*; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology*
chemical_list: D000906:Antibodies; D000939:Epitopes; D010455:Peptides; D011994:Recombinant Proteins