Abstract
Identifying the targets of antibody responses during infection is important for designing vaccines, developing diagnostic and prognostic tools, and understanding pathogenesis. We developed a novel deep sequence-coupled biopanning approach capable of identifying the protein epitopes of antibodies present in human polyclonal serum. Here, we report the adaptation of this approach for the identification of pathogen-specific epitopes recognized by antibodies elicited during acute infection. As a proof-of-principle, we applied this approach to assessing antibodies to Dengue virus (DENV). Using a panel of sera from patients with acute secondary DENV infection, we panned a DENV antigen fragment library displayed on the surface of bacteriophage MS2 virus-like particles and characterized the population of affinity-selected peptide epitopes by deep sequence analysis. Although there was considerable variation in the responses of individuals, we found several epitopes within the Envelope glycoprotein and Non-Structural Protein 1 that were commonly enriched. This report establishes a novel approach for characterizing pathogen-specific antibody responses in human sera, and has future utility in identifying novel diagnostic and vaccine targets.
Highlights
Knowing the targets of the antibodies that are elicited in response to natural infection is important for developing vaccines and new diagnostic tests
We recently described a strategy that uses peptide libraries displayed on the bacteriophage MS2 virus-like particle (MS2VLP) affinity selection platform and deep sequence analysis to identify epitopes targeted in serum from ovarian cancer patients [5]
In order to adapt the MS2-VLP affinity selection platform to the identification of linear epitopes recognized by sera of Dengue virus (DENV)-infected patients, we generated an antigen fragment library (AFL) of the DENV-3 polyprotein (Fig 1)
Summary
Knowing the targets of the antibodies that are elicited in response to natural infection is important for developing vaccines and new diagnostic tests. Our ability to comprehensively and quantitatively characterize the epitopes targeted by individual antibodies in a polyclonal population is limited. Recent efforts to couple deep-sequencing technologies with phage display-based biopanning provides an alternative and complementary strategy for characterizing epitopes targeted in complex polyclonal serum [1,2,3,4,5,6]. We recently described a strategy that uses peptide libraries displayed on the bacteriophage MS2 virus-like particle (MS2VLP) affinity selection platform and deep sequence analysis to identify epitopes targeted in serum from ovarian cancer patients [5]. We report the adaptation of this method to the PLOS ONE | DOI:10.1371/journal.pone.0171511. We report the adaptation of this method to the PLOS ONE | DOI:10.1371/journal.pone.0171511 February 2, 2017
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