Epitopes of Naturally Acquired and Vaccine‐Induced Anti‐Ebola Virus Glycoprotein Antibodies in Single Amino Acid Resolution

Heidepriem, Jasmin; Krähling, Verena; Dahlke, Christine; Wolf, Timo; Klein, Florian; Addo, Marylyn M.; Becker, Stephan; Loeffler, Felix F.

doi:10.1002/biot.202000069

Cited by 11 publications

(17 citation statements)

References 41 publications

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“…This comparison showed that we could identify the same IgG epitopes as the commercial reference (Table S8, Supporting Information), except for one minor signal (Figure 5d,e). Additionally, we found four previously reported epitopes, [ 34,35 ] as well as one to our knowledge unreported epitope (AA 430–448), which highlights that we can produce fully combinatorial high quality peptide microarrays. Subsequently, we synthesized microarrays of the complete Ebola virus proteome in single AA resolution (4805 individual peptides) on separate 8.18 × 13.36 mm 2 and 4.54 × 10.70 mm 2 areas to perform a comprehensive epitope study of the Ebola virus disease survivor (Figures S40–S47 and Tables S8–S15, Supporting Information).…”

Section: Resultssupporting

confidence: 52%

Automated Laser‐Transfer Synthesis of High‐Density Microarrays for Infectious Disease Screening

et al. 2022

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Laser‐induced forward transfer (LIFT) is a rapid laser‐patterning technique for high‐throughput combinatorial synthesis directly on glass slides. A lack of automation and precision limits LIFT applications to simple proof‐of‐concept syntheses of fewer than 100 compounds. Here, an automated synthesis instrument is reported that combines laser transfer and robotics for parallel synthesis in a microarray format with up to 10 000 individual reactions cm−2. An optimized pipeline for amide bond formation is the basis for preparing complex peptide microarrays with thousands of different sequences in high yield with high reproducibility. The resulting peptide arrays are of higher quality than commercial peptide arrays. More than 4800 15‐residue peptides resembling the entire Ebola virus proteome on a microarray are synthesized to study the antibody response of an Ebola virus infection survivor. Known and unknown epitopes that serve now as a basis for Ebola diagnostic development are identified. The versatility and precision of the synthesizer is demonstrated by in situ synthesis of fluorescent molecules via Schiff base reaction and multi‐step patterning of precisely definable amounts of fluorophores. This automated laser transfer synthesis approach opens new avenues for high‐throughput chemical synthesis and biological screening.

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Section: Resultssupporting

confidence: 52%

Automated Laser‐Transfer Synthesis of High‐Density Microarrays for Infectious Disease Screening

et al. 2022

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“…Out of the 89 peptide microarray serological studies included IgG was the most invested antibody type followed by IgM. The IgG response shows a more specific binding pattern (less noise) than the IgM response, which reflects the higher specificity of IgGs [ 21 ]. Two studies by Mishara et al [ 22 ] and Tokarz et al [ 23 ] investigated IgG and IgM profiles in cerebrospinal fluid.…”

Section: Resultsmentioning

confidence: 99%

Scoping review of the applications of peptide microarrays on the fight against human infections

et al. 2022

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Introduction This scoping review explores the use of peptide microarrays in the fight against infectious diseases. The research domains explored included the use of peptide microarrays in the mapping of linear B-cell and T cell epitopes, antimicrobial peptide discovery, immunosignature characterisation and disease immunodiagnostics. This review also provides a short overview of peptide microarray synthesis. Methods Electronic databases were systematically searched to identify relevant studies. The review was conducted using the Joanna Briggs Institute methodology for scoping reviews and data charting was performed using a predefined form. The results were reported by narrative synthesis in line with the Preferred Reporting Items for Systematic reviews and Meta-Analyses extension for Scoping Reviews guidelines. Results Ninety-five articles from 103 studies were included in the final data charting process. The majority (92. 0%) of the articles were published during 2010–2020 and were mostly from Europe (44.2%) and North America (34.7%). The findings were from the investigation of viral (45.6%), bacterial (32. 0%), parasitic (23.3%) and fungal (2. 0%) infections. Out of the serological studies, IgG was the most reported antibody type followed by IgM. The largest portion of the studies (77.7%) were related to mapping B-cell linear epitopes, 5.8% were on diagnostics, 5.8% reported on immunosignature characterisation and 8.7% reported on viral and bacterial cell binding assays. Two studies reported on T-cell epitope profiling. Conclusion The most important application of peptide microarrays was found to be B-cell epitope mapping or antibody profiling to identify diagnostic and vaccine targets. Immunosignatures identified by random peptide microarrays were found to be applied in the diagnosis of infections and interrogation of vaccine responses. The analysis of the interactions of random peptide microarrays with bacterial and viral cells using binding assays enabled the identification of antimicrobial peptides. Peptide microarray arrays were also used for T-cell linear epitope mapping which may provide more information for the design of peptide-based vaccines and for the development of diagnostic reagents.

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“…A number of immunodominant epitopes have been identified within the MLD and the glycan cap in animal studies, vaccine clinical trials and human survivors [49][50][51]. Interestingly, rather consistent antibody signatures have been identified in the less glycosylated GP2, whereas there is little correlation of antibody epitopes in GP1 containing mucin-like domain and glycan cap [52]. In non-human primates, vaccination approaches have generated the most immunodominant epitopes in the MLD, where class switched antibodies were also mapped to [51].…”

Section: Discussionmentioning

confidence: 99%

Isoform-specific O-glycosylation dictates Ebola virus infectivity

Bagdonaite

Abdurahman

Mirandola

et al. 2022

Preprint

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Ebola virus glycoprotein is one of the most heavily O-glycosylated viral envelope glycoproteins. The glycoprotein possesses a large mucin-like domain responsible for the cytopathic effect on infected cells, yet its structure or potential role in early entry events is poorly defined. To understand the importance of elongated O-glycans and the individual O-glycosylation sites for viral infectivity, we performed a comprehensive characterization of site-specific glycosylation governed by the three key GalNAc-transferases, GalNAc-T1, -T2, and -T3, initiating O-glycan biosynthesis. Using TMT isobaric labelling we performed quantitative differential O-glycoproteomics on proteins produced in wild type HEK293 cells and cell lines ablated for each of the three GalNAc-Ts, as well as compared it to patterns on wild type virus-like particles. In total we found 38 and 41 O-glycosites on virus like particle-derived and recombinant GP, respectively, with well correlated sites and site-specific structures. Examination of the isoform-specific glycosylation demonstrate selective initiation of a subset of O-glycosites by each enzyme, with GalNAc-T1 having the largest contribution. We next demonstrate that O-linked glycan truncation and perturbed initiation retarded the production of viral particles and decreased infectivity of progeny virus. This work represents a comprehensive site-specific analysis of EBOV GP and sheds light on differential regulation of EBOV GP glycosylation initiated by host GalNAc-Ts. Together with the effect on viral propagation it opens prospective avenues for tailored intervention approaches and means for modulating immunogen O-glycan density.

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Epitopes of Naturally Acquired and Vaccine‐Induced Anti‐Ebola Virus Glycoprotein Antibodies in Single Amino Acid Resolution

Cited by 11 publications

References 41 publications

Automated Laser‐Transfer Synthesis of High‐Density Microarrays for Infectious Disease Screening

Automated Laser‐Transfer Synthesis of High‐Density Microarrays for Infectious Disease Screening

Scoping review of the applications of peptide microarrays on the fight against human infections

Isoform-specific O-glycosylation dictates Ebola virus infectivity

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