Development and validation of an ELISA for quantitation of Bovine Viral Diarrhea Virus antigen in the critical stages of vaccine production

A. Pecora, M. S.Perez Aguirreburualde, D. Rodriguez, C. Seki, M. S. Levy, D. Bochoeyer, M. J. Dus Santos, A. Wigdorovitz

Research output: Contribution to journalArticlepeer-review

11 Scopus citations

Abstract

Bovine Viral Diarrhea Virus (BVDV) is the causative agent of a worldwide disease. The virus infects bovines of all ages, causing reproductive problems and contaminating biological products of high commercial value. The large-scale production of BVDV vaccines presents the challenge of processing antigenic proteins that are highly susceptible to the processing environment. Potency testing requires the immunization of cattle in order to determine the neutralizing antibodies titers induced by the vaccine. An alternative to the in vivo test is an in vitro measurement of key viral antigens. This paper describes the development and validation of a sandwich-type indirect ELISA that is able to detect and quantify BVDV E2 glycoprotein in live and inactivated BVDV. Validation parameters such as repeatability, intermediate precision, and reproducibility indicated that the developed ELISA constitutes an advanced tool for evaluating the BVDV antigen throughout manufacturing and vaccine release testing.

Original languageEnglish (US)
Pages (from-to)170-178
Number of pages9
JournalJournal of Virological Methods
Volume162
Issue number1-2
DOIs
StatePublished - Jan 2009

Bibliographical note

Funding Information:
This work was financially supported by Biogénesis-Bagó S.A. Argentina. The authors want to thank Ayelén Sammarruco for technical laboratory assistance and to INTA's Adventicious Virus Laboratory, Herpesvirus Laboratory and Gastroenteric Virus Laboratory for their collaboration in the inter-laboratory trial. The authors are especially grateful to Dr. Viviana Parreño (INTA) for her collaboration in cattle experiments, to Lucia Makek (INTA) for her help in statistical analysis and to Tarit K. Mukhopadhyay (University College London) for his help in editing.

Keywords

  • BVDV
  • ELISA
  • Validation

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