Enhancing Production of Recombinant Proteins from Mammalian Cells
Name
MEBCS012.pdf
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231.81 KB
Format
Adobe PDF
Checksum (MD5)
7ef6b5237c1e2f5af578422859991170
Author(s)
Wong, Victor V.T.
Wong, Niki S.C.
Tan, Hong-Kiat
Wang, Daniel I.C.
Yap, Miranda G.S.
Date Issued
January 2003
Series/Report no.
Molecular Engineering of Biological and Chemical Systems (MEBCS);
Abstract
The bio-manufacturing of recombinant proteins from mammalian cell cultures requires robust processes that can maximize protein yield while ensuring the efficacy of these proteins as human therapeutics. Recognizing that the challenge of improving protein yield and quality can be met through various approaches, this paper presents three strategies currently being developed in our group. A method for rapidly selecting subpopulations of cells with high production characteristics is proposed. This method combines the efficiency of green fluorescent protein/fluorescence-activated cell sorting (GFP/FACS)–based screening with homologous recombination to generate and select high-producing subclones. Next, the development of chemically defined, protein-free media for enhancing monoclonal antibody production is described. Analysis of culture media effects on the genome-wide transcriptional program of the cell is presented as a means to optimize the culture media and identify potential targets for genetic manipulation. Finally, we propose a method for increasing the extent of intracellular sialylation by improving the transport of CMP-sialic acid into the trans-Golgi. This is hypothesized to increase the sialic acid availability, and may enhance the degree of sialylation in the glycoprotein product.
Subjects
clonal selection
glycosylation
mammalian cell culture
media design
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