Optimization of Downstream Processing of a Monoclonal Antibody
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Authors
Yee, Yvonne
McAndrews, Kathleen
Hutson, Nikita
Emmons, Matthew
Issue Date
2010
Type
Thesis
Language
en_US
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Abstract
For the past few years, Genentech has worked closely with the Chemical
Engineering Department at the University of Nevada Reno on the antibody
separation processes for Lucentis. Lucentis is a humanized anti vascular endothelial
growth factor (VEGF) monoclonal antibody fragment used to treat macular
degeneration. VEGF promotes blood vessel growth in the wet form of macular
degeneration which leads to bleeding within the macula and causes blindness.
The purpose of this project was to study and design the antibody separation
processes which include the homogenizer, centrifugation, chromatography, and
membrane separation. The design of the process is to manufacture pure Lucentis
which is a fragment antigen binding that is expressed in genetically engineered
E.coli. Lucentis “like” material was available to run chromatography and separation
experiments on.
The design began with 1,000 L of fermented E. coli culture that contains
approximately 5 g/L of antibody and 20-25% of solid content (cells and cellular
debris). The final production concentration should be between 10-30 g/L with
99% purity.
The cells are first lysed using a homogenizer. The lysed cells are then centrifuged,
where any cell debris is removed. Next, chromatography steps are used to separate
the product. The final step of the process is tangential flow filtration. This removes
any additional impurities and prepares Lucentis for clinical use.
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In Copyright(All Rights Reserved)