title: Towards a peptide-based suspension array for the detection of pestivirus antibodies in swine.
abstract: Classical swine fever (CSF) is a highly contagious and lethal disease in swine. Serological tests for the diagnosis of CSF need not only to detect antibodies against CSFV, but also need to differentiate these from antibodies against other pestiviruses. To investigate the possibilities of specific peptide-based serology, various synthetic peptides that represent a well-described linear epitope of the CSFV E2 protein (TAVSPTTLR) were used to test the viability of a peptide-based suspension array for the detection of antibodies against pestiviruses in swine. The results show that N-terminally biotinylated peptides can bind to avidin conjugated beads, and function in detection of the corresponding monoclonal antibody WH303. There are indications that the length of the spacer between epitope and biotin affect the efficiency of the peptide-antibody interaction. A protocol was established that enables probing for antibodies in porcine sera, where neutravidin-blocking of serum and the use of empty control beads for normalization was crucial. With a set of porcine sera with antibodies against various pestiviruses, the proof of concept of a peptide-based suspension array for specific detection of antibodies against pestiviruses in porcine sera was demonstrated.
authors.affiliation: Central Veterinary Institute, Part of Wageningen UR, P.O. Box 65, 8200 AB Lelystad, The Netherlands. Electronic address: fimme.vanderwal@wur.nl.
authors.forename: Fimme J
authors.initials: FJ
authors.lastname: van der Wal
authors.affiliation: Central Veterinary Institute, Part of Wageningen UR, P.O. Box 65, 8200 AB Lelystad, The Netherlands. Electronic address: tinka.jelsma@wur.nl.
authors.forename: Tinka
authors.initials: T
authors.lastname: Jelsma
authors.affiliation: Central Veterinary Institute, Part of Wageningen UR, P.O. Box 65, 8200 AB Lelystad, The Netherlands. Electronic address: helmi.fijten@wur.nl.
authors.forename: Helmi
authors.initials: H
authors.lastname: Fijten
authors.affiliation: Central Veterinary Institute, Part of Wageningen UR, P.O. Box 65, 8200 AB Lelystad, The Netherlands. Electronic address: rene.achterberg@wur.nl.
authors.forename: René P
authors.initials: RP
authors.lastname: Achterberg
authors.affiliation: Central Veterinary Institute, Part of Wageningen UR, P.O. Box 65, 8200 AB Lelystad, The Netherlands. Electronic address: willie.loeffen@wur.nl.
authors.forename: Willie L A
authors.initials: WLA
authors.lastname: Loeffen
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000097:blood*; D000939:Epitopes / Q000276:immunology; D010455:Peptides / Q000276:immunology*; D010576:Pestivirus / Q000276:immunology*; D018182:Pestivirus Infections / Q000175:diagnosis* / Q000276:immunology / Q000821:virology; D040081:Protein Array Analysis / Q000379:methods*; D013552:Swine; D014759:Viral Envelope Proteins / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000939:Epitopes; D010455:Peptides; D014759:Viral Envelope Proteins
keywords: Classical swine fever virus; Peptide; Serum; Suspension array