Pathogen-specific deep sequence-coupled biopanning: A method for surveying human antibody responses.
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.
Department of Molecular Genetics and Microbiology, University of New Mexico School of Medicine, MSC08 4660, 1 University of New Mexico, Albuquerque, NM, United States of America.
Kathryn M
http://orcid.org/0000-0002-1428-8899
KM
Frietze
Gorgas Memorial Institute for Health Studies, Ave. Justo Arosemena y Calle 35, Panamá, Panamá.
Juan M
JM
Pascale
Gorgas Memorial Institute for Health Studies, Ave. Justo Arosemena y Calle 35, Panamá, Panamá.
Brechla
B
Moreno
Department of Molecular Genetics and Microbiology, University of New Mexico School of Medicine, MSC08 4660, 1 University of New Mexico, Albuquerque, NM, United States of America.
Bryce
B
Chackerian
Department of Molecular Genetics and Microbiology, University of New Mexico School of Medicine, MSC08 4660, 1 University of New Mexico, Albuquerque, NM, United States of America.
David S
DS
Peabody
D000914:Antibodies, Viral / Q000276:immunology; D000917:Antibody Formation / Q000502:physiology*; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology; D003715:Dengue / Q000276:immunology; D003716:Dengue Virus / Q000235:genetics / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000235:genetics / Q000276:immunology*; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D007239:Infections / Q000276:immunology*; D017909:Levivirus / Q000276:immunology; D010446:Peptide Fragments / Q000235:genetics / Q000276:immunology; D019151:Peptide Library
D000914:Antibodies, Viral; D000956:Antigens, Viral; D000939:Epitopes; D007074:Immunoglobulin G; D010446:Peptide Fragments; D019151:Peptide Library