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Purification of white spot syndrome virus by iodixanol density gradient centrifugation

(2013) JOURNAL OF FISH DISEASES. 36(10). p.841-851
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Abstract
Up to now, only a few brief procedures for purifying white spot syndrome virus (WSSV) have been described. They were mainly based on sucrose, NaBr and CsCl density gradient centrifugation. This work describes for the first time the purification of WSSV through iodixanol density gradients, using virus isolated from infected tissues and haemolymph of Penaeus vannamei (Boone). The purification from tissues included a concentration step by centrifugation (2.5 h at 60 000 g) onto a 50% iodixanol cushion and a purification step by centrifugation (3 h at 80 000 g) through a discontinuous iodixanol gradient (phosphate-buffered saline, 5%, 10%, 15% and 20%). The purification from infected haemolymph enclosed a dialysis step with a membrane of 1 000 kDa (18 h) and a purification step through the earlier iodixanol gradient. The gradients were collected in fractions and analysed. The number of particles, infectivity titre (in vivo), total protein and viral protein content were evaluated. The purification from infected tissues gave WSSV suspensions with a very high infectivity and an acceptable purity, while virus purified from haemolymph had a high infectivity and a very high purity. Additionally, it was observed that WSSV has an unusually low buoyant density and that it is very sensitive to high external pressures.
Keywords
white spot syndrome virus buoyant density, Penaeus vannamei, white spot syndrome virus infectivity, iodixanol, white spot syndrome virus purification, PENAEUS-VANNAMEI JUVENILES, FREE LITOPENAEUS-VANNAMEI, WATER TEMPERATURE 33-DEGREES-C, MONOCLONAL-ANTIBODIES, ORAL ROUTES, WSSV, PROTEIN, SHRIMP, INFECTION, CRAYFISH

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Chicago
Pereira Dantas Da Rocha Lima, João José, Mathias Corteel, Maria Cornelissen, Peter Bossier, Patrick Sorgeloos, and Hans Nauwynck. 2013. “Purification of White Spot Syndrome Virus by Iodixanol Density Gradient Centrifugation.” Journal of Fish Diseases 36 (10): 841–851.
APA
Pereira Dantas Da Rocha Lima, J. J., Corteel, M., Cornelissen, M., Bossier, P., Sorgeloos, P., & Nauwynck, H. (2013). Purification of white spot syndrome virus by iodixanol density gradient centrifugation. JOURNAL OF FISH DISEASES, 36(10), 841–851.
Vancouver
1.
Pereira Dantas Da Rocha Lima JJ, Corteel M, Cornelissen M, Bossier P, Sorgeloos P, Nauwynck H. Purification of white spot syndrome virus by iodixanol density gradient centrifugation. JOURNAL OF FISH DISEASES. 2013;36(10):841–51.
MLA
Pereira Dantas Da Rocha Lima, João José, Mathias Corteel, Maria Cornelissen, et al. “Purification of White Spot Syndrome Virus by Iodixanol Density Gradient Centrifugation.” JOURNAL OF FISH DISEASES 36.10 (2013): 841–851. Print.
@article{3192024,
  abstract     = {Up to now, only a few brief procedures for purifying white spot syndrome virus (WSSV) have been described. They were mainly based on sucrose, NaBr and CsCl density gradient centrifugation. This work describes for the first time the purification of WSSV through iodixanol density gradients, using virus isolated from infected tissues and haemolymph of Penaeus vannamei (Boone). The purification from tissues included a concentration step by centrifugation (2.5 h at 60 000 g) onto a 50\% iodixanol cushion and a purification step by centrifugation (3 h at 80 000 g) through a discontinuous iodixanol gradient (phosphate-buffered saline, 5\%, 10\%, 15\% and 20\%). The purification from infected haemolymph enclosed a dialysis step with a membrane of 1 000 kDa (18 h) and a purification step through the earlier iodixanol gradient. The gradients were collected in fractions and analysed. The number of particles, infectivity titre (in vivo), total protein and viral protein content were evaluated. The purification from infected tissues gave WSSV suspensions with a very high infectivity and an acceptable purity, while virus purified from haemolymph had a high infectivity and a very high purity. Additionally, it was observed that WSSV has an unusually low buoyant density and that it is very sensitive to high external pressures.},
  author       = {Pereira Dantas Da Rocha Lima, Jo{\~a}o Jos{\'e} and Corteel, Mathias and Cornelissen, Maria and Bossier, Peter and Sorgeloos, Patrick and Nauwynck, Hans},
  issn         = {0140-7775},
  journal      = {JOURNAL OF FISH DISEASES},
  keyword      = {white spot syndrome virus buoyant density,Penaeus vannamei,white spot syndrome virus infectivity,iodixanol,white spot syndrome virus purification,PENAEUS-VANNAMEI JUVENILES,FREE LITOPENAEUS-VANNAMEI,WATER TEMPERATURE 33-DEGREES-C,MONOCLONAL-ANTIBODIES,ORAL ROUTES,WSSV,PROTEIN,SHRIMP,INFECTION,CRAYFISH},
  language     = {eng},
  number       = {10},
  pages        = {841--851},
  title        = {Purification of white spot syndrome virus by iodixanol density gradient centrifugation},
  url          = {http://dx.doi.org/10.1111/jfd.12082},
  volume       = {36},
  year         = {2013},
}

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