Isolation and characterization of Newcastle disease virus from biological fluids using column chromatography

Abstract

Newcastle disease virus (NDV), belonging to the species avian orthoavulavirus 1, genus Orthoavulavirus, and family Paramyxoviridae, is responsible for Newcastle disease in poultry and other avian species. It has shown significant potential as an oncolytic virus and as a vector for vaccine delivery. NDV from infected biological serum is usually isolated or purified using density gradient ultracentrifugation. However, it has many disadvantages, including the fact that it is time consuming and can process only a limited quantity of sample at one time. In our study, native agarose gel electrophoresis and dynamic light scattering (DLS) analysis showed that NDV carried a net negative surface charge. Thus, we purified the virus using a HiTrap Q Sepharose Fast Flow anion exchange column with salt elution. Hemagglutination assay and plaque assay showed that the procedure yielded high‐purity NDV particles with a recovery of more than 80%, and the process was fast and simple. The purity of the virus was confirmed using sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE) and Western blot analysis. The hydrodynamic volume and ‘dry state’ diameter of the purified NDV were analyzed using dynamic light scattering and transmission electron microscopy and were to be in the range of 200–300 nm. The viruses did not exhibit any deviation from their known physical properties. The genome of the virus was also detected by amplifying a 423‐bp region using reverse transcription‐polymerase chain reaction. Our study confirmed that NDV could be effectively purified using an anion exchange column. In addition, the procedure could be easily upscaled or downscaled based on the experimental requirements.

Ecological and Molecular Genetic Characterization of Newcastle Disease Virus

“…Экологическая характеристика вируса болезни Ньюкасла. Вирус болезни Ньюкасла инфицированными птицами распространяется в течение 1-2 недель после заражения с фекалиями и выделениями из дыхательных путей [12], а также через яйцо вылупившемуся цыпленку [13]. Вирус присутствует во всех частях тушки и способен в холодильнике сохраняться в течение многих месяцев [9].…”

Enhanced Downstream Processing for a Cell-Based Avian Influenza (H5N1) Vaccine

“…Nevertheless, this approach falls short in removing host cell residues, specifically residual protein and residual DNA, which are crucial impurities in cell culture. In virus purification, gel filtration (GF) chromatography is widely employed to purify most viruses due to the distinct differences in molecular weight, density, and charge properties between virus molecules and impurities [ 15 ]. GF chromatography is significantly faster, more consistent, and easier to automate compared to the conventional method [ 16 ].…”

Characterization of Chicken-Derived Genotype VII Newcastle Disease Virus Isolates from Northwest China

“…NDV has strong infectivity, spreads rapidly, is associated with high morbidity and fatality, and causes multiple diseases in birds. Based on mean death time (MDT), intracerebral pathogenicity index (ICPI), and intravenous pathogenicity index (IVPI), NDV strains have been characterized as velogenic, mesogenic, and lentogenic ( Cox and Plemper, 2017 ; Gao et al, 2019 ; Neog et al, 2023 ). Associated clinical signs include high fever, dyspnea, dysentery, nervous disorders, and bleeding of mucous and serous membranes, causing serious financial losses in the poultry industry ( Jia et al, 2018 ).…”