Reverse Engineering of Vaccine Antigens Using High Throughput Sequencing-enhanced mRNA Display
Autor: | Philip R. Krause, Hongying Duan, Benjamin W. Krause, Nini Guo, Marian E. Major, Alla Kachko |
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Rok vydání: | 2015 |
Předmět: |
Viral Hepatitis Vaccines
medicine.drug_class lcsh:Medicine Context (language use) Hepacivirus Protein Engineering Monoclonal antibody General Biochemistry Genetics and Molecular Biology Epitope Antibodies Monoclonal Murine-Derived Epitopes Mice Antigen Vaccine reverse-engineering medicine Animals mRNA display RNA Messenger Peptide library Antigens Viral lcsh:R5-920 biology Mimotope lcsh:R General Medicine Hepatitis C Antibodies Antibodies Neutralizing Virology biology.protein Original Article Antibody lcsh:Medicine (General) |
Zdroj: | EBioMedicine EBioMedicine, Vol 2, Iss 8, Pp 859-867 (2015) |
ISSN: | 2352-3964 |
DOI: | 10.1016/j.ebiom.2015.06.021 |
Popis: | Vaccine reverse engineering is emerging as an important approach to vaccine antigen identification, recently focusing mainly on structural characterization of interactions between neutralizing monoclonal antibodies (mAbs) and antigens. Using mAbs that bind unknown antigen structures, we sought to probe the intrinsic features of antibody antigen-binding sites with a high complexity peptide library, aiming to identify conformationally optimized mimotope antigens that capture mAb-specific epitopes. Using a high throughput sequencing-enhanced messenger ribonucleic acid (mRNA) display approach, we identified high affinity binding peptides for a hepatitis C virus neutralizing mAb. Immunization with the selected peptides induced neutralizing activity similar to that of the original mAb. Antibodies elicited by the most commonly selected peptides were predominantly against specific epitopes. Thus, using mRNA display to interrogate mAbs permits high resolution identification of functional peptide antigens that direct targeted immune responses, supporting its use in vaccine reverse engineering for pathogens against which potent neutralizing mAbs are available. Research in Context We used a large number of randomly produced small proteins (“peptides”) to identify peptides containing specific protein sequences that bind efficiently to an antibody that can prevent hepatitis C virus infection in cell culture. After the identified peptides were injected into mice, the mice produced their own antibodies with characteristics similar to the original antibody. This approach can provide previously unavailable information about antibody binding and could also be useful in developing new vaccines. Highlights • mRNA-display/high throughput sequencing identified high affinity peptide binders to monoclonal antibody (mAb). • The profile of selected peptides characterized the binding specificity of the selection mAb. • Immune responses induced by selected peptides were neutralizing and epitope-focused, mimicking the selection mAb. |
Databáze: | OpenAIRE |
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