Towards a peptide-based suspension array for the detection of pestivirus antibodies in swine

Fimme J. van der Wal*, Tinka Jelsma, Helmi Fijten, René P. Achterberg, Willie L.A. Loeffen

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

4 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)15-20
JournalJournal of Virological Methods
Volume235
DOIs
Publication statusPublished - 2016

Fingerprint

Pestivirus
Suspensions
Swine
Peptides
Antibodies
Classical Swine Fever
Serum
Epitopes
Swine Diseases
Avidin
Serologic Tests
Serology
Biotin
Monoclonal Antibodies

Keywords

  • Classical swine fever virus
  • Peptide
  • Serum
  • Suspension array

Cite this

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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.",
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author = "{van der Wal}, {Fimme J.} and Tinka Jelsma and Helmi Fijten and Achterberg, {Ren{\'e} P.} and Loeffen, {Willie L.A.}",
year = "2016",
doi = "10.1016/j.jviromet.2016.04.022",
language = "English",
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pages = "15--20",
journal = "Journal of Virological Methods",
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Towards a peptide-based suspension array for the detection of pestivirus antibodies in swine. / van der Wal, Fimme J.; Jelsma, Tinka; Fijten, Helmi; Achterberg, René P.; Loeffen, Willie L.A.

In: Journal of Virological Methods, Vol. 235, 2016, p. 15-20.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Towards a peptide-based suspension array for the detection of pestivirus antibodies in swine

AU - van der Wal, Fimme J.

AU - Jelsma, Tinka

AU - Fijten, Helmi

AU - Achterberg, René P.

AU - Loeffen, Willie L.A.

PY - 2016

Y1 - 2016

N2 - 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.

AB - 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.

KW - Classical swine fever virus

KW - Peptide

KW - Serum

KW - Suspension array

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DO - 10.1016/j.jviromet.2016.04.022

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