Purification of poliovirus labeled with radiophosphorus

Taylor, Joyce; Graham, Angus

doi:10.1016/0042-6822(58)90096-5

Cited by 26 publications

(6 citation statements)

References 15 publications

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“…The catecholamine depletion observed after guanethidine ( 12.5 mg/kg) is slower however than aiter reserpine (4). Guanethidine, like reserpine, produces a decline in content of tissue nore-Ilinephrine in the rabbit and cat.…”

mentioning

confidence: 90%

A Simple Method for Concentration of Live and Formaldehyde-Inactivated Poliovirus

Grossowicz

Mercado

Goldblum

1960

Experimental Biology and Medicine

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mentioning

confidence: 90%

A Simple Method for Concentration of Live and Formaldehyde-Inactivated Poliovirus

Grossowicz

Mercado

Goldblum

1960

Experimental Biology and Medicine

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“…Although 32p has frequently been used to label animal viruses in their nucleic acid (4,5,11), 33P, which is now available from commercial sources in acceptable radioisotope purity, appears to offer several advantages over 32P as a radiotracer for phosphorus in viruses and other biological materials (8). Both 32P and 33P decay by beta emission to produce stable sulfur atoms.…”

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confidence: 99%

Preparation of Poliovirus Labeled with Phosphorus-33

Sobsey

Lavigne

Cooper

1972

Applied Microbiology

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Phosphorus-33 (33P), a weak (0.25 Mev) beta-emitting isotope of phosphorus with a half-life of 25 days, has been used to label poliovirus in cell culture. HeLa cell monolayers were depleted of phosphate and then labeled by incubating at 37 C in a medium (LM) containing about 10 gCi of 33P as orthophosphate per ml. Labeled cells were infected at a high multiplicity with poliovirus type 1 and incubated for 8 hr in LM medium. Virus from infected cells was then concentrated and purified. Virus purity was confirmed by comparison of virus infectivity and radioactivity after CsCl density gradient centrifugation and by observing purified virus preparations with electron microscopy. With the method described, yields of about 1010 to 5 x 1010 plaque-forming units (PFU) of highly purified poliovirus with specific activities of about 3 x 10-4 to 10-3disintegrations per min per PFU have been obtained from 1.5 x 108 to 3.0 x 108 HeLa cells.

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“…phosphate (5,11), and poliovirus and enteroviruses by precipitation with cations (6,7,10), by chromatography on cellulose ion-exchange columns (8,18), and by precipitation with methanol with further purification by elution, ultracentrifugation, and enzymatic treatment (9,17).…”

mentioning

confidence: 99%

“…In the third procedure, the supernatant after tissuecell removal was cooled to -1 C in a dry ice and water mixture, and precooled absolute methanol (-10C) was added in a ratio of 1:5 (alcohol to virus supematant). The virus-alcohol mixture was held at -10 C ovemight or approximately 16 to 18 hr, after which the mixture was centrifuged for 30 min at 6,000 X g in the SS-34 rotor of a Servall RC-2 centrifuge operating at -10 C. The supematant was decanted, and the precipitate was suspended in an equal volume of peptone-supplemented medium 199 with 10% bovine serum added.…”

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confidence: 99%

Concentration of Rift Valley Fever and Chikungunya Viruses by Precipitation

Klein¹,

Mahlandt²,

Cockey³

et al. 1970

Applied Microbiology

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Simple and efficient methods for concentrating Rift Valley fever (RVF) virus and chikungunya (CHIK) virus are described. Ammonium sulfate, potassium sulfate, or alcohol was used as a precipitating agent and the precipitate was resuspended to volumes suitable for further processing and purification. The methods permitted concentration of live RVF virus and CHIK virus about 100-fold with negligible losses of virus. RVF virus retained a high level of infectivity with potassium aluminum sulfate and alcohol, but CHIK virus retained a higher infectivity level with ammonium sulfate than with potassium aluminum sulfate. The data indicate that serum plays an important role in the concentration of both viruses, at least when the sulfate methods are used. Certain viruses have been concentrated by adsorption to salts and molecular gels, e.g., polioviruses on aluminum hydroxide gels (14, 16) and on calcium and aluminum phosphate gel (19), fowl plaque influenza, Newcastle, and mumps viruses on calcium phosphate (3, 15) or aluminum

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Purification of poliovirus labeled with radiophosphorus

Cited by 26 publications

References 15 publications

A Simple Method for Concentration of Live and Formaldehyde-Inactivated Poliovirus

A Simple Method for Concentration of Live and Formaldehyde-Inactivated Poliovirus

Preparation of Poliovirus Labeled with Phosphorus-33

Concentration of Rift Valley Fever and Chikungunya Viruses by Precipitation

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