The immunome of soy bean allergy: Comprehensive identification and characterization of epitopes

Abstract

Summary Background The precise mapping of multiple antibody epitopes recognized by patients’ sera allows a more detailed and differentiated understanding of immunological diseases. It may lead to the development of novel therapies and diagnostic tools. Objective Mapping soy bean specific epitopes relevant for soy bean allergy patients and persons sensitized to soy bean, and analysis of their IgE/IgG binding spectrum. Methods Identification of epitopes using sera, applying an optimized peptide phage display library followed by next‐generation sequencing, specially designed in silico data analysis and subsequent peptide microarray analysis. Results We were able to identify more than 400 potential epitope motifs in soy bean proteins. More than 60% of them have not yet been described as potential epitopes. Eighty‐three peptides, representing the 42 most frequently found epitope candidates, were validated by microarray analysis using 50 sera from people who have been tested positive in skin prick test (SPT). Of these peptides, 56 were bound by antibodies, 55 by serum IgE, 43 by serum IgG and 30 by both. Person‐specific epitope patterns were found for each individual and protein. Conclusions For individuals with clinical symptoms, epitope resolved analyses reveal a high prevalence of IgE binding to a few soy bean specific epitopes. Evaluation of individual immune profiles of patients with soy bean sensitization allows the identification of peptides that do facilitate studying individual IgE/IgG epitope binding patterns. This enables discrimination of sensitization from disease, such assay test has the potential to replace SPT assays

Clinical utility of microarray B‐cell epitope mapping in food allergies: A systematic review

“…Five epitope‐mapping studies, one methodological study, and one diagnostic study did not perform statistical analysis or did not provide information about it.…”

“…Recently, Kern et al identify more than 400 potential epitope motifs in soybean proteins applying peptide phage display technology . The epitope candidates were validated using two types of peptide microarrays, the classical fixed‐length overlapping peptide microarray, and a microarray printed with variable‐length peptide epitopes identified by the phage display approach.…”

Identification of Seasonal Variations of Antibodies against PR-10-Specific Epitopes Can Be Improved Using Peptide-Phage Display

“…For this proof of concept study, 4 patients were sufficient. Larger patient cohorts will be needed to identify season-relevant epitopes because epitope recognition patterns of different patients are highly heterogeneous [7,9,10]. Cor a 1…”

“…Current methods such as peptide phage display combined with next generation sequencing [6,[9][10][11][12][13] and overlapping peptide microarrays [7,8] have severe limitations regarding the small number of identified epitopes or the inability to identify conformational epitopes, respectively. We recently reported the identification of more than 400 potential epitope motifs from soya allergens directly by antibodies from sensitized patients' sera using a peptide phage display platform followed by epitope validation through peptide microarrays [10]. This method is based on a unique statistical motif-oriented analysis approach of peptide phage display data [10,12].…”

IgE Epitope Profiling for Allergy Diagnosis and Therapy – Parallel Analysis of a Multitude of Potential Linear Epitopes Using a High Throughput Screening Platform

“…By screening the phage library with the desired ligand(s) potential binders can be isolated, propagated and sequenced in order to reveal the introduced amino acid sequence motif which interacted with the ligand(s). In allergology this technique has been used to study IgE-allergen interaction, either by cloning single chain antibody genes into the phage ( 17 – 19 ) and offering the gene products to a given allergen, or vice versa by offering a defined IgE reactivity to a pool of peptide-presenting phages created by cloning random oligonucleotides into the permissive site of the scaffold protein gene ( 20 – 23 ). The latter variant – defined IgE reactivity versus a broad peptide landscape – is more common as it may yield information about the epitope recognized by the IgE in question.…”