PREPARATION OF A STABLE NONINFECTIVE GROUP-SPECIFIC ADENOVIRUS DIAGNOSTIC ANTIGEN

1964 ◽  
Vol 10 (5) ◽  
pp. 747-751
Author(s):  
John R. Polley ◽  
Muriel M. Guerin
Keyword(s):  
Tissue Culture ◽  
Culture Fluid ◽  
Copper Hydroxide ◽  
Diagnostic Antigen ◽  
Sintered Glass ◽  
Antigen Present ◽  
Glass Filter ◽  
Sintered Glass Filter ◽  
Serological Reactivity ◽  
Infected Cells

In the preparation of a group-specific diagnostic antigen for adenoviruses, various procedures for the concentration and purification of the antigen present in the tissue culture fluid of specifically infected cells were investigated. It was found possible to prepare an antigen of increased potency and specificity by passage of the fluid through an ultrafine sintered glass filter, followed by coprecipitation of the antigen with copper hydroxide. This antigen was rendered noninfective by treatment with 0.01% formaldehyde at pH 7 at 37 °C for 4 days or by heating at 56 °C at pH 5.5 for 3 hours. After destruction of the infectivity the antigen was stabilized for storage by lyophilization. This antigen, after such treatment, still retained its serological reactivity.

Comparative Study of Micro Infrared Techniques

1970 ◽  
Vol 53 (3) ◽  
pp. 599-603
Author(s):  
Wilson L Brannon
Keyword(s):  
Infrared Spectroscopy ◽  
Comparative Study ◽  
Silica Gel ◽  
Isolation And Identification ◽  
Sintered Glass ◽  
Isolation Techniques ◽  
Glass Filter ◽  
Sintered Glass Filter ◽  
Beam Center ◽  
Infrared Techniques

Abstract A comparative study was made of several techniques used in the isolation and identification of microgram quantities of drugs by infrared spectroscopy. The isolation techniques include preparative GLC and TLC. In the case of TLC, silica gel interference was nearly eliminated by using a sintered glass filter and vacuum. The identifications were enhanced by concentrating the sample in the infrared beam center. The sample was centered either with the aid of a cardboard silhouette or punched out or drilled indentations in the disk.

scholarly journals POLARIZATION STUDIES IN COLLODION MEMBRANES AND IN SYNTHETIC PROTEIN-LIPOID MEMBRANES

1937 ◽  
Vol 20 (5) ◽  
pp. 695-710 ◽  
Author(s):  
Mona Spiegel-Adolf
Keyword(s):  
Olive Oil ◽  
Glass System ◽  
Limited Time ◽  
Low Resistance ◽  
Sintered Glass ◽  
Glass Filter ◽  
Sintered Glass Filter ◽  
Synthetic Protein ◽  
Polarization Studies

1. Collodion membranes of high polarizability and low resistance can be obtained either by addition of certain ether-soluble substances such as phosphatides, olive oil, mastix, and gum benzoin, to the collodion or by drying collodion membranes for a limited time under pressure. 2. The permeability of membranes of different polarization has been measured by means of conductivity methods. 3. Sintered glass filter plates of Jena glass crucibles on which proteins and lipoids have been adsorbed show polarization. It could be shown that some narcotics which react with lecithin cause an increase in polarization of the protein-lipoid-glass system. Substitutions of the protein but not of the lipoid were possible, without causing a decrease in the polarizability of the membranes.

Preparation of a purified adenovirus diagnostic antigen by gel filtration

1969 ◽  
Vol 15 (10) ◽  
pp. 1167-1171 ◽  
Author(s):  
John R. Polley ◽  
T. S. Webb
Keyword(s):  
Tissue Culture ◽  
Gamma Irradiation ◽  
Gel Filtration ◽  
Culture Fluid ◽  
Pressure Filtration ◽  
Routine Production ◽  
Diagnostic Antigen ◽  
Sepharose 4B ◽  
Molecular Sieving

Gel filtration has been investigated as a method for purifying adenovirus antigens in tissue culture fluid. It was found that by molecular sieving through the agarose Sepharose 4B, most of the infectivity, hemagglutinins, and nonspecific complement-fixing (CF) activity could be separated from a second fraction which contained some hemagglutinin but only a low titer of viable virus and from a third fraction which contained the group-specific complement-fixing antigen but was relatively free of hemagglutinin, viable virus, and nonspecific reactants.For the routine production of a noninfective, purified group-specific diagnostic antigen for adenovirus, the bulk infected fluid is inactivated by heating at 56 °C for 3 hours at pH 5.5 or by gamma irradiation with 2.5 megarads, concentrated about 20-fold by pervaporation or by pressure filtration, purified by gel filtration using Sepharose 4B, concentrated to desired potency, and then lyophilized for stable storage.

scholarly journals Dengue Virus Type 1 Nonstructural Glycoprotein NS1 Is Secreted from Mammalian Cells as a Soluble Hexamer in a Glycosylation-Dependent Fashion

1999 ◽  
Vol 73 (7) ◽  
pp. 6104-6110 ◽  
Author(s):  
Marie Flamand ◽  
Françoise Megret ◽  
Magali Mathieu ◽  
Jean Lepault ◽  
Félix A. Rey ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Virus Type ◽  
Mammalian Cells ◽  
Culture Fluid ◽  
Vero Cells ◽  
Soluble Form ◽  
Complex Type ◽  
Infected Insect ◽  
Infected Cells

ABSTRACT Nonstructural glycoprotein NS1, specified by dengue virus type 1 (Den-1), is secreted from infected green monkey kidney (Vero) cells in a major soluble form characterized by biochemical and biophysical means as a unique hexameric species. This noncovalently bound oligomer is formed by three dimeric subunits and has a molecular mass of 310 kDa and a Stokes radius of 64.4 Å. During protein export, one of the two oligosaccharides of NS1 is processed into an endo-β-N-acetylglucosaminidase F-resistant complex-type sugar while the other remains of the polymannose type, protected in the dimeric subunit from the action of maturation enzymes. Complete processing of the complex-type sugar appears to be required for efficient release of soluble NS1 into the culture fluid of infected cells, as suggested by the repressive effects of the N-glycan processing inhibitors swainsonine and deoxymannojyrimicin. These results, together with observations related to the absence of secretion of NS1 from Den-infected insect cells, suggest that maturation and secretion of hexameric NS1 depend on the glycosylation status of the host cell.

scholarly journals BIOCHEMICAL STUDY OF CELLULAR ANTIGEN-ANTIBODY REACTION IN TISSUE CULTURE

1960 ◽  
Vol 112 (2) ◽  
pp. 249-255 ◽  
Author(s):  
Akira Tokuda ◽  
Hideo Hayashi ◽  
Kinishiro Matsuba
Keyword(s):  
Tissue Culture ◽  
Protease Activity ◽  
Biochemical Study ◽  
Culture Fluid ◽  
Partial Purification ◽  
Trichloracetic Acid ◽  
Heat Stable ◽  
Antibody Reaction ◽  
Antigen Antibody ◽  
Cellular Antigen

Decrease in the protease activity of the culture fluid observed at later stages of the antigen-antibody reaction is believed to be due to the release of an inhibitor by the cells. The inhibitor was submitted to partial purification: it is heat-stable, non-precipitated by trichloracetic acid and non-dialyzable. It inhibits certain cellular and tissue proteases and papain but is inactive against trypsin. It is suggested that the balance between protease and anti-protease released may determine the intensity, extent, and duration of certain sensitization phenomena.

scholarly journals Immunosuppressive activity of a subline of the mouse EL-4 lymphoma. Evidence for minute virus of mice causing the inhibition.

1976 ◽  
Vol 143 (1) ◽  
pp. 187-205 ◽  
Author(s):  
G D Bonnard ◽  
E K Manders ◽  
D A Campbell ◽  
R B Herberman ◽  
M J Collins
Keyword(s):  
Tissue Culture ◽  
Culture Fluid ◽  
Inhibitory Factor ◽  
Minute Virus Of Mice ◽  
G Cells ◽  
Culture Cells ◽  
Tissue Culture Cells ◽  
Lymphocyte Cultures ◽  
Minute Virus ◽  
Kilham Rat Virus

Filtered culture fluids from the early in vitro passages of a subline of the C57BL/6 mouse EL-4 lymphoma, EL-4(G-), were strongly inhibitory for BABL/c vs. C57BL/6 mixed lymphocyte cultures (MLC). The inhibitory activity could be preserved by storage at -75 degrees C or 4 degrees C, thus allowing its further characterization. The inhibitory factor was particulate (nondialyzable, sedimentable at 100,000 g for 1 h), very small (recovered after 0.10 mum filtration), sensitive to UV irradiation, but heat stable (56 degrees C, 1 h) and resistant to chloroform. It was infectious, since later, noninhibitory passages of EL-4(G-) tissue culture cells became strongly inhibitory upon inoculation with the culture fluid. This data was consistent with the inhibitory factor being an infectious virus. Virus analysis by mouse antibody production tests revealed that viruses were indeed present in EL-4(G-) ascites cells and in the culture fluid, and not in a late passage of EL-4(G-) tissue culture cells which were not inhibitory. Neutralization of the inhibitory factor was achieved by pretreatment with ascitic fluid or with the sera raised against those (EL-4(G-)-derived materials which contained viruses. Mouse reference immune sera against minute virus of mice (MVM) completely neutralized the inhibitory factor in the culture fluid or in EL-4(G-) ascites cells. Two prototype MVM strains, and one Kilham rat virus preparation, did not inhibit the mouse MLC. Thus, the possibility exists that a variant of MVM, or an unidentified virus, has been grown and selected for in EL-4(G-) cells and recognized, due to its immunosuppressive characteristics. In any event, immunosuppression by EL-4(G-) cells was not mediated by the tumor cells, their metabolic products, or associated endogenous type C viruses, but by an exogenous virus, most likely a variant MVM with immunosuppressive characteristics. This adds weight to a parallel observation from our laboratory on the immunosuppressive effects of Kilham rat virus in rat lymphocyte cultures.

Plasma protein recovery from spent tissue culture fluid

1991 ◽  
Vol 13 (4) ◽  
pp. 261-264 ◽  
Author(s):  
P. K. Ng ◽  
C. Figueroa ◽  
G. Mitra
Keyword(s):  
Tissue Culture ◽  
Plasma Protein ◽  
Culture Fluid ◽  
Protein Recovery

Stability of visna virus in infectious tissue culture fluid

1961 ◽  
Vol 10 (4) ◽  
pp. 501-509 ◽  
Author(s):  
Halld�r Thormar
Keyword(s):  
Tissue Culture ◽  
Culture Fluid ◽  
Visna Virus
Sign in / Sign up

Export Citation Format

Share Document