title: Validation study to evaluate the reproducibility of a candidate in vitro potency assay of newcastle disease vaccines and to establish the suitability of a candidate biological reference preparation.
abstract: A quantification assay for the Haemagglutinin-Neuraminidase (HN) protein of Newcastle Disease Virus (NDV) has been developed at CIDC-Lelystad as a candidate in vitro potency test for inactivated Newcastle disease (ND) vaccines. In studies performed at CIDC-Lelystad, a high correlation was demonstrated between the results of this candidate in vitro potency assay and the results of the serological potency assay (European Pharmacopoeia monograph 0870; test A). Furthermore, a high correlation between the serological data (Haemagglutination Inhibition-antibody titres) and clinical protection after challenge was demonstrated. Correlation between in vivo and in vitro potency assays was confirmed in a collaborative pre-validation study. In the pre-validation study three Official Medicines Control Laboratories (OMCLs) determined both the NDV-HN antigen content and the in vivo potency (vaccination-serology and vaccination-challenge) of 6 vaccine batches. The conclusion of the pre-validation study was that a large-scale collaborative study should be organised to validate the in vitro method and the suitability of the reference preparation. This report describes the outcome of this study. In brief, 14 laboratories (8 OMCLs and 6 vaccine manufacturers) determined the NDV-HN antigen content of 9 different vaccines in 3 independent tests. The vaccine batches were produced by 5 different manufacturers and represent a quantitative range of ND antigen content. One vaccine batch with insufficient potency and one poultry vaccine not containing NDV were included. Statistical evaluation of the results indicated that the antigen content could be determined with high precision. A good repeatability as well as reproducibility was found. Furthermore all laboratories found a similar ranking of the vaccines, based on the antigen content. Comparison of the antigen content and the in vivo potency of a series of vaccines with relatively low potencies indicated that a threshold relative antigen level of 7.0 antigen units per dose would discriminate between vaccine batches with sufficient and insufficient potency. An in vitro assay with this threshold level for antigen content did not result in any false positive results and only a limited number of false negative results in the BSP055 study. We conclude that the in vitro measurement of the antigen content of inactivated ND-vaccines with the proposed method is a reliable alternative potency assay that could be included as a new method in monograph 0870 on ND-vaccines.
authors.affiliation: CIDC-Lelystad, PO Box 2004, 8203 AA Lelystad, The Netherlands.
authors.forename: I
authors.initials: I
authors.lastname: Claassen
authors.forename: R
authors.initials: R
authors.lastname: Maas
authors.forename: H
authors.initials: H
authors.lastname: Oei
authors.forename: A
authors.initials: A
authors.lastname: Daas
authors.forename: C
authors.initials: C
authors.lastname: Milne
mesh_terms: D000818:Animals; D000956:Antigens, Viral / Q000276:immunology; D001717:Birds; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D009521:Newcastle Disease / Q000276:immunology*; D009522:Newcastle disease virus / Q000276:immunology*; D012015:Reference Standards; D015203:Reproducibility of Results; D014765:Viral Vaccines / Q000592:standards*
chemical_list: D000956:Antigens, Viral; D014765:Viral Vaccines