A critical avenue for the improvement of production of biotherapeutics in mammalian cells is to increase the stability of clonal cell lines. This study investigates the molecular basis for the onset of clonal heterogeneity in different recombinant protein (h-IgG) secreting CHO clones by characterizing specific mechanisms of transgene silencing, such as genetic drift, epigenetic modifications, and biochemical pathway perturbations. Using the Cell Xpress™ software module on the LEAP (Laser-Enabled Analysis and Processing) instrument platform, we assessed the relative levels of heterogeneity, as determined by specific cell productivity, in clonal CHO recombinant cell lines. Subpopulations of specific clones were generated by isolating cells from the highest 25% of secretors based on Cell Xpress™ data. These populations were analyzed via transcriptional profiling, copy number analysis, and epigenetic characterization to identify defining properties of cells that maintain or lose short-term production stability.

01 Jan 2010·Cells and Culture

An Evaluation of the Intrinsic IgG Production Capabilities of Different Chinese Hamster Ovary Parental Cell Lines

Author: Nan Lin ; Kevin J. Kayser ; Genova A. Richardson ; Daniel W. Allison ; Matthew V. Caple

To better characterize differences in expression and secretion capacity, we quantitatively analyzed production of Green Fluorescent Protein (GFP) and secretion of recombinant human IgG in transiently transfected CHOK1SV, ECACC K1 and CHO DG44 parental cell lines. By analyzing these production trends in a transient transfection system, we were able to compare recombinant protein production and secretion between the different CHO parental cell lines independent of integration site effect.

01 Jan 2009·Cell Engineering

Cell Xpress™ Applications in Development and Characterization of Biopharmaceutical Recombinant Protein Producing Cell Lines

Author: Genova A. Richardson ; Kevin J. Kayser ; Jennifer R. Cresswell ; Nan Lin

A review.The biopharmaceutical industry is focused on the development of quality processes for producing therapeutic proteins and monoclonal antibodies in mammalian cells.The first step in the process development workflow is cell line development.This chapter focuses on applications developed on the LEAP (Laser-Enabled Anal. and Processing) instrument to expedite the cell line development process.Applications reviewed include expression optimization, high throughput single cell clone isolation and cell population characterization.

100 Deals associated with Sigma-Aldrich Co. Ltd.

100 Translational Medicine associated with Sigma-Aldrich Co. Ltd.

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