Enhancing Production of Recombinant Proteins from Mammalian Cells
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 a...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | en_US |
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2003
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| Online Access: | http://hdl.handle.net/1721.1/3782 |
| _version_ | 1811087409963597824 |
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| author | Wong, Victor V.T. Wong, Niki S.C. Tan, Hong-Kiat Wang, Daniel I.C. Yap, Miranda G.S. |
| author_facet | Wong, Victor V.T. Wong, Niki S.C. Tan, Hong-Kiat Wang, Daniel I.C. Yap, Miranda G.S. |
| author_sort | Wong, Victor V.T. |
| collection | MIT |
| description | 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. |
| first_indexed | 2024-09-23T13:45:39Z |
| format | Article |
| id | mit-1721.1/3782 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T13:45:39Z |
| publishDate | 2003 |
| record_format | dspace |
| spelling | mit-1721.1/37822019-04-12T08:15:25Z Enhancing Production of Recombinant Proteins from Mammalian Cells Wong, Victor V.T. Wong, Niki S.C. Tan, Hong-Kiat Wang, Daniel I.C. Yap, Miranda G.S. clonal selection glycosylation mammalian cell culture media design 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. Singapore-MIT Alliance (SMA) 2003-12-08T15:20:00Z 2003-12-08T15:20:00Z 2003-01 Article http://hdl.handle.net/1721.1/3782 en_US Molecular Engineering of Biological and Chemical Systems (MEBCS); 237369 bytes application/pdf application/pdf |
| spellingShingle | clonal selection glycosylation mammalian cell culture media design Wong, Victor V.T. Wong, Niki S.C. Tan, Hong-Kiat Wang, Daniel I.C. Yap, Miranda G.S. Enhancing Production of Recombinant Proteins from Mammalian Cells |
| title | Enhancing Production of Recombinant Proteins from Mammalian Cells |
| title_full | Enhancing Production of Recombinant Proteins from Mammalian Cells |
| title_fullStr | Enhancing Production of Recombinant Proteins from Mammalian Cells |
| title_full_unstemmed | Enhancing Production of Recombinant Proteins from Mammalian Cells |
| title_short | Enhancing Production of Recombinant Proteins from Mammalian Cells |
| title_sort | enhancing production of recombinant proteins from mammalian cells |
| topic | clonal selection glycosylation mammalian cell culture media design |
| url | http://hdl.handle.net/1721.1/3782 |
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