Evolving serodiagnostics by rationally designed peptide arrays: the Burkholderia paradigm in Cystic Fibrosis

Abstract

Efficient diagnosis of emerging and novel bacterial infections is fundamental to guide decisions on therapeutic treatments. Here, we engineered a novel rational strategy to design peptide microarray platforms, which combines structural and genomic analyses to predict the binding interfaces between diverse protein antigens and antibodies against Burkholderia cepacia complex infections present in the sera of Cystic Fibrosis (CF) patients. The predicted binding interfaces on the antigens are synthesized in the form of isolated peptides and chemically optimized for controlled orientation on the surface. Our platform displays multiple Burkholderia-related epitopes and is shown to diagnose infected individuals even in presence of superinfections caused by other prevalent CF pathogens, with limited cost and time requirements. Moreover, our data point out that the specific patterns determined by combined probe responses might provide a characterization of Burkholderia infections even at the subtype level (genomovars). The method is general and immediately applicable to other bacteria.

Layer-by-layer deposition of functional click polymers for microarray applications

“…This antigen is part of the OMPA (Outer Membrane Protein A) family of proteins, a group of genetically related proteins responsible for bacterial survival. Previous studies have validated peptide OMPA3 as a diagnostic probe for Burkholderia infections in subjects affected by cystic fibrosis [26] and investigated the effectiveness of different peptide-to-surface conjugation strategies [17]. We found that all the click chemoselective reactions that were evaluated, i.e.…”

Targeting Difficult Protein-Protein Interactions with Plain and General Computational Approaches

“…In general, it is tempting to suggest that the possibility to predict the parts of a protein (antigens) endowed with antibody recognition/binding properties and the demonstration of their reactivity in the form of isolated peptides can open up new venues for diagnosis and treatment. In the case of diagnostics, for instance, multiple predicted binding sequences can be displayed on microarrays for medium-high throughput analysis of their interaction profiles: in a notable instance, predicted peptides were optimized for oriented display on microarray plates and proved to be efficient in the rapid diagnosis of Burkholderia infections in cystic fibrosis (CF) patients [ 61 ]. To mimic conformational epitopes, oriented and spatially controlled co-immobilization of predicted epitope sequences that are spatially proximal in the Zika virus NS1 protein, showed the ability to cooperatively interact to provide enhanced immunoreactivity with respect to single linear epitopes [ 62 ].…”

Postgenomic Approaches and Bioinformatics Tools to Advance the Development of Vaccines against Bacteria of the Burkholderia cepacia Complex

“…Epitope identification is expensive and laborious, requiring experimental screening of large arrays of potential epitope candidates. However, recent works of epitope mapping of Burkholderia strains have been performed using the peptide microarray approach [ 59 , 60 , 61 ]. This technique relies on the use of libraries of synthesized linear peptides spotted on a single chip and screening for antigenicity using patients’ antisera.…”