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A Rapid Method for the Capture and Purification of a Protease Sensitive Protein

by Christopher W. Kemp, PhD
Volume 1, Issue 2 (Summer 2002)

The baculovirus expression vector system (BEVS) is one method utilized for the production of recombinant proteins, and typically works without significant difficulties. However, some proteins are produced in insoluble forms, and degradation can occur. This article will focus on this degradation issue, and present a method to stabilize a protease-sensitive protein that has been produced at the 40-liter scale...

Citation:
Kemp CW. A Rapid Method for the Capture and Purification of a Protease Sensitive Protein. BioProcess J, 2002; 1(2): 36-39.

 
Risk Analysis of Raw Materials Used in Mammalian Cell Culture Media

by Trish Benton and Thomas C. Thomas
Volume 1, Issue 2 (Summer 2002)

Growth media for mammalian cell culture are complex mixtures of raw materials that include amino acids, vitamins, inorganic salts and a wide variety of other components. The risk of infectious agent transmission, when some of these components are derived directly from animals, is a major concern in the biopharmaceutical industry, and is being actively addressed. However, the risk associated with the use of indirectly, or secondarily, derived animal components is less recognized and addressed. We have developed a classification system to define the contact level that a cell culture medium component has had with animal-derived materials. This classification system has increased the accuracy and reliability of the information we are able to obtain from raw material manufacturers, and is being used as part of a risk assessment analysis for a serum/protein-free media we are moving from development into manufacturing...

Citation:
Benton T, Thomas TC. Risk Analysis of Raw Materials Used in Mammalian Cell Culture Media. BioProcess J, 2002; 1(2): 40-42.

 
Large-Scale Production of Active Serine-Threonine Kinases of the MAPK Pathway Using the Baculovirus System

by Steven L. Nguyen and Wai-Ping Fung-Leung, PhD
Volume 1, Issue 2 (Summer 2002)

Serine-threonine kinases of the Mitogen Activated Protein Kinase (MAPK) pathway represent potential drug targets for a wide range of diseases. As part of an effort to understand the biology of the pathways, several human serine-threonine MAPKs were produced. Optimization and modification of current methodologies used in the baculovirus expression system resulted in the generation of large amounts of active MAPKs. Compounds found to inhibit the MAPKs in vitro, subsequently showed activity in cell-based assays and animal models. The processes to be discussed were developed to yield large quantities of three active human serine-threonine MAPKs by the baculovirus expression system...

Citation:
Nguyen SL, Fung-Leung W-P. Large-Scale Production of Active Serine-Threonine Kinases of the MAPK Pathway Using the Baculovirus System. BioProcess J, 2002; 1(2): 43-46.

 
Regulatory Aspects of Recombinant Protein Products by Baculovirus Expression Systems

by Daniel Galbraith, PhD
Volume 1, Issue 2 (Summer 2002)

The baculovirus expression system promises to revolutionize the production of recombinant proteins for use as clinical products. The technology is robust, efficient, and low-cost when compared to other cell based systems. The technique may also present an advantage in producing safer products versus the equivalent materials made with mammalian cells. Proteins can be produced in insect cells without animal supplements such as fetal calf serum. In the current climate of concerns over Bovine Spongiform Encephalopathies, and bovine viral risks, this method offers a significant safety, as well as cost advantage, over other production methods...

Citation:
Galbraith D. Regulatory Aspects of Recombinant Protein Products by Baculovirus Expression Systems. BioProcess J, 2002; 1(2): 47-51.

 
Production of a p55gag Particle Vaccine Using the Baculovirus Expression Vector System Technology

by Penny L. Post, PhD and Manon M.J. Cox
Volume 1, Issue 2 (Summer 2002)

Globally, an estimated 36 million people are living with HIV, and some 20 million people have already died of AIDS. Today, there is still no HIV vaccine available. HIV virus-like particles are an attractive vaccine candidate due to their ability to induce both antibody and cytotoxic T-lymphocyte responses. In this article, we describe the development of a production process for an HIV particle vaccine, HIV-1 p55 (gag). The gag precursor protein (p55) is sufficient for assembly and cellular release of retrovirus-like particles. We expressed the p55 gag protein using the BEVS technology in Spodoptera frugiperda expresSF+ cells...

Citation:
Post PL, Cox MMJ. Production of a p55gag Particle Vaccine Using the Baculovirus Expression Vector System Technology. BioProcess J, 2002; 1(2): 52-59.

 
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