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Adenovirus Type 5 (Ad5) Chromatographic Purification Process at the 20 L Scale

by Normand Arcand, Alice Bernier, Julia Transfiguracion, Danielle Jacob, Helene Coelho, and Amine Kamen, PhD
Volume 2, Issue 1 (January/February 2003)

Recombinant adenovirus are attractive as vectors for gene therapy because: they exhibit wide tissue tropism and high transduction efficiency; adenovirus cultures can reach high specific titers (10^10 VP/mL), and; their use in the treatment of cancer and other serious diseases is valuable. A primary mode of adenovirus purification continues to be CsCl density gradient centrifugation...

Citation:
Arcand N, Bernier A, Transfiguracion J, Jacob D, Coelho H, Kamen A. Adenovirus Type 5 (Ad5) Chromatographic Purification Process at the 20 L Scale. BioProcess J, 2003; 2(1): 72-75.

 
Methods for Detection and Evaluation of Replication Competent Adenovirus (RCA)

by Dongling Ma, Amanda Newman, William T. Lucas, Renee N. Meloro, Lisa Rudderow, Joseph V. Hughes, and Garry B. Takle
Volume 1, Issue 3 (Fall 2002)

In the past, researchers developing gene therapy applications used replication-defective human Adenovirus 5 (Ad5) as a vector for delivering DNA sequences, almost exclusively. Ad5 vectors are typically rendered replication defective by the deletion of E1a gene sequences. A complementing cell line containing the E1a gene makes it possible to produce Ad5 vectors in large scale. Of the various cell lines that have been constructed for the purpose of high-titer Ad5 production, HEK293 cells and PER.C6 cells are the most widely used...

Citation:
Ma D, Newman A, Lucas WT, Meloro RN, Rudderow L, Hughes JV, Takle GB. Methods for Detection and Evaluation of Replication Competent Adenovirus (RCA). BioProcess J, 2002; 1(3): 26-30.

 
Production Criteria in the Development of a K562-Based Vaccine Platform

by R. Ripley, N. Mahesh, W. Press, J-S. Lee, E. Aguilar-Cordova, and E.J. Beecham
Volume 1, Issue 3 (Fall 2002)

The K562 cell line is a human myelogenous leukemic cell which has been used by several groups, including ours, as a vehicle for cell-based vaccines and immuno-gene therapies. The attractiveness of K562 cells is the ease with which they can be cultured, plus the fact that they express very low levels of MHC proteins. Low MHC expression facilitates the use of these cells in patients with different MHC backgrounds, and it may improve the in vivo survival of the cells by delaying immune rejection. Based largely on these properties, we have been developing the K562 cell line as a universal platform for expressing cytokines, tumor antigens, and other immuno-modulating proteins...

Citation:
Ripley R, Mahesh N, Press W, Lee J-S, Aguilar-Cordova E, Beecham EJ. Production Criteria in the Development of a K562-Based Vaccine Platform. BioProcess J, 2002; 1(3): 31-35.

 
Chemical Cell Lysis for Large-Scale Adenoviral Vector Production

by Mark I. Fitchmun, PhD, C. Lanio, B. Grimm, E. Prokopenko, J. Zhou, and S. Josephs
Volume 1, Issue 3 (Fall 2002)

Production of non-enveloped viruses generally requires a cell lysis procedure to liberate mature particles trapped within their host cells. The standard bench-scale practice of using freeze/thaw cycles is simple and effective, but heat transfer limitations restrict the technique to relatively small applications. Here we show that a ten-minute treatment with a dilute mixture of polysorbate-80 and tri-butyl phosphate effectively liberates adenovirus from host cells...

Citation:
Fitchmun MI, Lanio C, Grimm B, Propopenko E, Zhou J, Josephs S. Chemical Cell Lysis for Large-Scale Adenoviral Vector Production. BioProcess J, 2002; 1(3): 36-40.

 
FDA Perspectives on the Use of the Adenovirus Reference Material

by Stephanie Simek, PhD, Andrew Byrnes, PhD, and Steven Bauer, PhD
Volume 1, Issue 3 (Fall 2002)

As development proceeds for adenoviral vectors in gene transfer clinical trials, it becomes increasingly important that these products demonstrate a good safety profile, and thereby build confidence in those who must make decisions about risk/benefit ratios, dose escalation, and efficacy. Currently, safety and efficacy are based predominantly upon the analysis of data generated by non-standardized methods, resulting in inconsistent values being reported for virus titer and particle counts...

Citation:
Simek S, Byrnes A, Bauer S. FDA Perspectives on the Use of the Adenovirus Reference Material. BioProcess J, 2002; 1(3): 40-42.

 
A Statistical Analysis of Adenovirus Reference Material Assay Results

by Janice D. Callahan, PhD
Volume 1, Issue 3 (Fall 2002)

The Adenovirus Reference Material (ARM) was developed under the guidance of the Adenovirus Reference Material Working Group (ARMWG) and the U.S. Food and Drug Administration (FDA), and was made possible through the donation of services and supplies by a large number of laboratories and institutions from the United States, Canada, France, The Netherlands, Germany, and the United Kingdom. The purpose of the ARM is to provide a reference material for use in validating assays and internal standards for adenoviral particle concentration and infectious titer. The NIH Recombinant DNA Advisory Committee recommended the development of such a reference-testing agent in their report issued January 2002. The ARM consists of purified wild type 5 Adenovirus as described by ATCC's catalog number VR-5...

Citation:
Callahan JD. A Statistical Analysis of Adenovirus Reference Material Assay Results. BioProcess J, 2002; 1(3): 43-47.

 
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