Large-scale isolation and partial purification of type C RNA viruses on hydroxyapatite

Smith, R G; Lee, Sin A.

doi:10.1016/0003-2697(78)90340-8

Cited by 12 publications

(7 citation statements)

References 16 publications

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“…Virus particles were concentrated and purified from supernatant fluids of induced cultures by adsorption and release from hydroxyapatite (HAP) at phosphate ion concentrations of 0.15 M and 0.4 M, respectively (Smith & Lee, 1978). These virus preparations were centrifuged at 100000 g for 60 rain, and the pellets were resuspended in a total volume of 9.6 ml of 30 ~ Percoll (Pharmacia) and 0.25 M-sucrose in 0.01 M-Tris-HC1 pH 7-4.…”

Section: Methodsmentioning

confidence: 99%

Production of Virions with Retrovirus Morphology by Human Embryonal Carcinoma Cells in vitro

Bronson

Saxinger

Ritzi

et al. 1984

Journal of General Virology

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SUMMARYEmbryonal carcinoma (EC) cell lines established from human testicular germ-cell tumours produce, at low frequency, virions morphologically identical to type C retroviruses that have been observed by other workers in human placental tissues. The virus particles are formed while budding from the cell surface, and their numbers are increased by inducing the EC ceils with 5-iodo-2'-deoxyuridine and dexamethasone. Assays for RNA-dependent DNA polymerase (reverse transcriptase) associated with purified virions suggested a low level of activity. In addition, another type of virus is occasionally produced by induced cells of three EC lines. These particles also form during the process of budding from the cell surface, but they have surface projections (spikes). Extracellular spiked virions frequently are pleomorphic, with a condensed, eccentric nucleoid, and thus morphologically resemble type B retroviruses. No virions of either type were detected with or without induction in cultures of differentiated EC cells or in cultures of yolk sac carcinoma or teratoma cells, both of which are considered malignant but differentiated derivatives of EC cells. The lack of virion production by these differentiated cells suggests developmental regulation of virus replication.

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Section: Methodsmentioning

confidence: 99%

Production of Virions with Retrovirus Morphology by Human Embryonal Carcinoma Cells in vitro

Bronson

Saxinger

Ritzi

et al. 1984

Journal of General Virology

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“…Hydroxyapatite has been used for the adsorption of other replication competent human viruses (Tsuru et al, 1991). It has also been used for the purification of viral proteins of several C-type retroviruses (including murine leukaemia viruses) but the recovery of infectious virus was not studied (Smith et al, 1978). Also, the crystalline form of hydroxyapatite used for these studies does not lend itself to regeneration of the column material, in contrast to the ceramic hydroxyapatite described here.…”

Section: Introductionmentioning

confidence: 92%

Purification of a functional gene therapy vector derived from Moloney murine leukaemia virus using membrane filtration and ceramic hydroxyapatite chromatography

Kuiper

Sanches

Walford

et al. 2002

Biotech & Bioengineering

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The ability of membrane ultra- and diafiltration and two chromatography media, Matrex Cellufine Sulfate (Millipore) and Macro-Prep ceramic hydroxyapatite (Bio-Rad), to adsorb, elute, and purify gene therapy vectors based on Moloney murine leukaemia virus (MoMuLV) carrying the 4070A amphotropic envelope protein was studied. Membrane ultra- and diafiltration provided virus concentration up to 160-fold with an average recovery of infectious viruses of 77 +/- 14%. In batch experiments, Macro-Prep ceramic hydroxyapatite (type 2, particle size 40 microm) proved superior to Matrex Cellufine Sulfate for MoMuLV vector particle adsorption. Furthermore, functional vector particles could be eluted using phosphate buffer pH 6.8 (highest titres from >or=300 mM phosphate) from the Macro-Prep adsorbent, with higher specific titres (cfu/mg protein) than the starting material. Similar results were obtained when this ceramic hydroxyapatite was packed into a column and used in a liquid chromatography system. Recovery of transduction-competent virus was between 18 and 31% for column experiments and 32 and 46% for batch experiments.

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“…Because the primary human leukemic bonemarrow cells and most of the cell lines developed from them were not available for analysis, all the experiments described herein were performed using virus derived from the experimentally infected rabbit corneal cells (SIRC) (denoted SKA21) or human A204 cells (denoted A204V). The murine fibroblast line JLS-VIO producing MuLV-R, BALB/c clone A31, human A673 cells producing BALB virus-2 (BC177) (Aaronson and Stevznson,I973), the human rhabdomyosarcoina cell linc (A204), human lymphoid cells (NC37) producing the San Francisco strain of the gibbon ape leukemia virus GaLVsF (Snyder et a/., 1973), and marmoset tumor cells producing the woolly monkey fibrosarcoma virus (7IAPl/SiSV-SSAV) (Theilen et a/., 1971), were grown according to methods previously described (Smith and Lee, 1978). The baboon endogenous virus (M7) (Benveniste et a/., 1974), A204 cells infected with M7, the gibbon ape leukemia virus (GaLV) (Kawakami et al, 1972;Snyder et nl., 19731, and BALB virus-2 (BC169) (Aaronson and Stevenson, 1973) were obtained from the Resources and Logistics Segment, National Cancer Institute.…”

Section: Cells and Virusesmentioning

confidence: 99%

Characterization of a type‐C virus produced by co‐cultures of human leukemic bone‐marrow and fetal canine thymus cells

Smith

Nooter

Bentvelzen

et al. 1979

Intl Journal of Cancer

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The putative human helper virus SKA-21/A204V, isolated by Nooter et al. in 1977 from human leukemic bone-marrow cells following co-culture with normal fetal canine thymus cells, Cf2th, has been characterized with respect to its major viral core protein, reverse transcriptase, and nucleic acid sequences. The results of these analyses show that this virus is not distinguishable from the woolly monkey type-C virus, SSAV-1, by the techniques employed.

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Large-scale isolation and partial purification of type C RNA viruses on hydroxyapatite

Cited by 12 publications

References 16 publications

Production of Virions with Retrovirus Morphology by Human Embryonal Carcinoma Cells in vitro

Production of Virions with Retrovirus Morphology by Human Embryonal Carcinoma Cells in vitro

Purification of a functional gene therapy vector derived from Moloney murine leukaemia virus using membrane filtration and ceramic hydroxyapatite chromatography

Characterization of a type‐C virus produced by co‐cultures of human leukemic bone‐marrow and fetal canine thymus cells

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