Abstract
Background: Antibodies to microbes, or to autoantigens, are important markers of disease. Antibody detection (serology) can reveal both past and recent infections. There is a great need for development of rational ways of detecting and quantifying antibodies, both for humans and animals. Traditionally, serology using synthetic antigens covers linear epitopes using up to 30 amino acid peptides. Methods: We here report that peptides of 100 amino acids or longer (“megapeptides”), designed and synthesized for optimal serological performance, can successfully be used as detection antigens in a suspension multiplex immunoassay (SMIA). Megapeptides can quickly be created just from pathogen sequences. A combination of rational sequencing and bioinformatic routines for definition of diagnostically-relevant antigens can, thus, rapidly yield efficient serological diagnostic tools for an emerging infectious pathogen. Results: We designed megapeptides using bioinformatics and viral genome sequences. These long peptides were tested as antigens for the presence of antibodies in human serum to the filo-, herpes-, and polyoma virus families in a multiplex microarray system. All of these virus families contain recently discovered or emerging infectious viruses. Conclusion: Long synthetic peptides can be useful as serological diagnostic antigens, serving as biomarkers, in suspension microarrays.
Highlights
The frequent appearance of novel pathogens [1,2,3,4,5], and reemergence of those who became rare [6], emphasizes the need for an ability to generically detect them, and rapidly evolve diagnostic techniques that can enhance the response to such challenges
Weoverview evaluatedofifthe a 116 amino acidserosurveillance megapeptide derived from the extracellular surface glycoprotein of Zaire Ebola virus would function as an antigen in suspension multiplex immunoassay (SMIA)
We evaluated if a 116protein amino from acid megapeptide derived fromrelated the extracellular portion2)
Summary
The frequent appearance of novel pathogens [1,2,3,4,5], and reemergence of those who became rare [6], emphasizes the need for an ability to generically detect them, and rapidly evolve diagnostic techniques that can enhance the response to such challenges. SMIA is a cost-effective serological method where antigen and serum consumption can be minimized, its background is generally low, and the dynamic range wider than in ELISA Unlike the latter, it provides multiple results per analysis [7,8,9,10]. Methods: We here report that peptides of 100 amino acids or longer (“megapeptides”), designed and synthesized for optimal serological performance, can successfully be used as detection antigens in a suspension multiplex immunoassay (SMIA). Results: We designed megapeptides using bioinformatics and viral genome sequences These long peptides were tested as antigens for the presence of antibodies in human serum to the filo-, herpes-, and polyoma virus families in a multiplex microarray system. All of these virus families contain recently discovered or emerging infectious viruses
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