Electrophoretic Studies on Pr8 Influenza Virus

Miller, Gail Lorenz; Lauffer, Max A.; Stanley, W. M.

doi:10.1084/jem.80.6.549

“…Yes dsDNA [170,176] Influenza virus Orthomyxoviridae 80-120 (H2N2) A2/Singapore/57: 5.0 Pleomorphic, spherical Yes (-)ssRNA [177][178][179][180][181][182][183] (H3N1) A/Pol/L/71: 6.5-6.8 (H3N1) A/Pol/20/72: 6.5-6.8 (H3N1) A/Enfl/42/72: 6.5-6.8 (H3N1) A/Phil/2/82: 6.5-6.8 (H3N1) A/Pol/79/85: 6.5-6.8 (H1N1) A1/Lenigrad: 5.0 (H3N2) A3/ Lenigrad): 5.0 (H1N1) A/PR/8: 5.3…”

Section: Reviewmentioning

confidence: 99%

Downstream processing of cell culture-derived virus particles

Wolf

¹

,

Reichl

²

2011

Expert Review of Vaccines

View full text Add to dashboard Cite

Manufacturing of cell culture-derived virus particles for vaccination and gene therapy is a rapidly growing field in the biopharmaceutical industry. The process involves a number of complex tasks and unit operations ranging from selection of host cells and virus strains for the cultivation in bioreactors to the purification and formulation of the final product. For the majority of cell culture-derived products, efforts focused on maximization of bioreactor yields, whereas design and optimization of downstream processes were often neglected. Owing to this biased focus, downstream procedures today often constitute a bottleneck in various manufacturing processes and account for the majority of the overall production costs. For efficient production methods, particularly in sight of constantly increasing economic pressure within human healthcare systems, highly productive downstream schemes have to be developed. Here, we discuss unit operations and downstream trains to purify virus particles for use as vaccines and vectors for gene therapy.

show abstract

“…Friedenreich [10] carried out extensive studies of the phenomenon and demonstrated the enzymatic nature of the agent. In the ensuing years twenty-one cases have been reported which are believed by some to be the examples of the occurrence of this phe nomenon in vivo [18,11,1,2,9,14,19,15,12,25,22,27,20,13]. The validity of the relationship of these cases to the Thomsen-Friedenreich phenomenon will be discussed later.…”

Section: Discussionmentioning

confidence: 98%

The Purification of Antibody to PR8 Influenza A Virus

lsliker¹,

Strauss²

1959

Vox Sanguinis

0

View full text Add to dashboard Cite

The physico-chemical procedures at present available for the separation of proteins appear to possess inadequate resolving power for the isolation of specific antibodies from mixtures such as the γ-globulin fraction of human plasma [23, 3, 14]. The failure to achieve significant purification by these means is due presumably to the similarity of the various antibodies in solubility, size, shape, net charge and other physical chemical properties. A different approach takes advantage of the natural interaction of an antibody with its homologous antigen to separate the specific antibody from the mixture [8, 9, 26]. The complex is then isolated and dissociated. The recovery of antibody is facilitated by linking the antigen to a solid phase [2, 13]. Cellulose or ion exchange resins, which offer several means of forming insoluble complexes with the antigen, have proved particularly useful. The difficulty remains, however, of achieving satisfactory dissociation of antibodies from protein antigens. In such cases good yields of antibody can be obtained only by use of extremely acid or alkaline reactions. The present study has been directed primarily at overcoming this difficulty. Working on the hypothesis that the formation of the anti-gen-antibody complex results in part from electrostatic interaction between the molecules, the dissociating action of a synthetic polyelectrolyte has been studied. Polymethacrylic acid for instance has been shown to interact with proteins depending on the relative charges of the reactants [16, 21, 27]. Since PR8 influenza A virus and γ-globulin (containing the homologous antibody) bear opposite net charges between pH 5.4 and 6.5 (interisoelectric range) the possibility of a selective interaction of polymethacrylic acid with either antigen or antibody was investigated. Experiments were first performed to study the solubility of the total γ-globulin fraction (antibody) and PR8 influenza A virus (antigen) in the presence of polymethacrylic acid (PMA). Various procedures were then carried out to isolate the soluble virus antibody complex which was to be fractionated with PMA. The method was compared with the purification of antibody by means of antigens linked to ion exchange resins [13]. Although the PR8 influenza A virus and its homologous antibody comprised the test system, it is anticipated that this procedure should prove applicable to the purification of other antibodies, when a relatively pure antigen is available.

show abstract

“…Friedenreieh [lo] carried out extensive studies of the phenomenon and demonstrated the enzymatic nature of the agent. In the ensuing years twenty-one cases have been reported which are believed by some t o be the examples of the occurrence of this phenomenon in vivo [18,11,1,2,9,14,19,15,12,25,22,27,20,131. The validity of the relationship of these cases t o the Thomsen-Friedenreich phenomenon will be discussed later.…”

Section: Discussionmentioning

confidence: 99%

The Purification of Antibody to PR8 Influenza A Virus

Isliker

¹

,

Stuuss

²

1959

View full text Add to dashboard Cite

The physico-chemical procedures a t present available for the separation of proteins appear to possess inadequate resolving power for the isolation of specific antibodies from mixtures such as the y-globulin fraction of human plasma [23, 3, 141. The failure t o achieve significant purification by these means is due presumably to the similarity of the various antibodies in solubiIity, size, shape, net charge and other physical chemical properties. A different approach takes advantage of the natural interaction of an antibody with its homologous antigen t o separate the specific antibody from the mixture [8, 9, 261. The complex is then isolated and dissociated. The recovery of antibody is facilitated by linking the antigen to a solid phase [2, 131. Cellulose or ion exchange resins, which offer several means of forming insoluble complexes witd the antigen, have proved particularly useful. The difficulty remains, however, of achieving satisfactory dissociation of antibodies from protein antigens. In such cases good yields of antibody can be obtained only b y use of extremely acid or alkaline reactions.The present study has been directed primarily a t overcoming this difficulty. Working on the hypothesis that the formation of the antigen-antibody complex results in part from electrostatic interaction between the molecules, the dissociating action of a synthetic polyelectrolyte has been studied. Polymethacrylic acid for instance has been shown t o interact with proteins depending on the relative

show abstract

Electrophoretic Studies on Pr8 Influenza Virus

Cited by 49 publications

References 15 publications

Downstream processing of cell culture-derived virus particles

Downstream processing of cell culture-derived virus particles

The Purification of Antibody to PR8 Influenza A Virus

The Purification of Antibody to PR8 Influenza A Virus

Contact Info

Product

Resources

About