Purification and concentration of infectious koi herpesvirus using steric exclusion chromatography.
J Fish Dis
; 46(8): 873-886, 2023 Aug.
Article
em En
| MEDLINE
| ID: mdl-37227769
Koi herpesvirus (KHV) is the causative agent of a koi herpesvirus disease (KHVD) inducing high mortality rates in common carp and koi (Cyprinus carpio). No widespread effective vaccination strategy has been implemented yet, which is partly due to side effects of the immunized fish. In this study, we present an evaluation of the purification of infectious KHV from host cell protein and DNA, using the steric exclusion chromatography. The method is related to conventional polyethylene glycol (PEG) precipitation implemented in a chromatographic set-up and has been applied for infectious virus particle purification with high recoveries and impurity removal. Here, we achieved a yield of up to 55% of infectious KHV by using 12% PEG (molecular weight of 6 kDa) at pH 7.0. The recoveries were higher when using chromatographic cellulose membranes with 3-5 µm pores in diameter instead of 1 µm. The losses were assumed to originate from dense KHV precipitates retained on the membranes. Additionally, the use of >0.6 M NaCl was shown to inactivate infectious KHV. In summary, we propose a first step towards a purification procedure for infectious KHV with a possible implementation in fish vaccine manufacturing.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Carpas
/
Doenças Transmissíveis
/
Infecções por Herpesviridae
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Doenças dos Peixes
/
Herpesviridae
Limite:
Animals
Idioma:
En
Revista:
J Fish Dis
Assunto da revista:
BIOLOGIA
/
MEDICINA VETERINARIA
Ano de publicação:
2023
Tipo de documento:
Article
País de afiliação:
Alemanha
País de publicação:
Reino Unido