Article | June 1, 2007
Proteomic Analysis Of CHO Cells During Recombinant Protein Production In High-Density Culture
•Proteomic Analysis Of CHO Cells During Recombinant Protein Production In High-Density Culture
A proteomics approach was used to study protein expression changes in a recombinant IgGproducing CHO cell line throughout a fed-batch culture. Protein expression profi les were analyzed by 2-dimensional gel electrophoresis at four time points representing mid exponential, late exponential, mid and late stationary phases of growth. Differentially expressed proteins were selected using Phoretix™ 2D Expression software and identifi ed using in-gel tryptic digest and LC-MS (MS). Molecules involved in cellular metabolism, protein synthesis and folding, antioxidation and cytoskeletal functions were identifi ed to be upregulated during the fed-batch bioreactor run. These studies provided insight in the intracellular mechanisms that correlate with high-level recombinant production and may shed some light on the signifi cance of protein folding and secretion in recombinant protein production. These results may also provide potential candidate genes for future cell line and metabolic engineering work.
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SOURCE: SAFC