کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
23884 43479 2011 6 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
High-throughput analysis of animal cell cultures using two-dimensional fluorometry
موضوعات مرتبط
مهندسی و علوم پایه مهندسی شیمی بیو مهندسی (مهندسی زیستی)
پیش نمایش صفحه اول مقاله
High-throughput analysis of animal cell cultures using two-dimensional fluorometry
چکیده انگلیسی

We report a new method which combines fluorescence spectroscopy at microtiter plate scale with multivariate statistical analysis for rapid and high-throughput analysis of secreted recombinant protein and viable cell growth in animal cell cultures. The potential of the method is demonstrated by application to cultures of three Chinese Hamster Ovary (CHO) cell clones with distinct IgG4 antibody yields. Supernatant samples collected throughout culture time were analysed by two-dimensional fluorometry; significant changes were observed in the regions of tryptophan, metabolic cofactors and vitamins. Partial least squares regression was then used to correlate the entire fluorescence map with measured concentrations of antibody and viable cells. For both target variables, a model was calibrated with representative data from the two less productive clones and validated with data from the best producer clone; this allowed viable cell density to be predicted for the validation clone with an average error of 10%; even better, the secreted antibody could be predicted with an average error of 7%, proving the predictive capacity of the model beyond the calibration region. All the main spectral regions were required to establish the best correlations for both targeted variables. In conclusion, this method effectively analyzes cellular productivity in 96-well plate format, shortening the time spent in early phases of bioprocess development.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Biotechnology - Volume 151, Issue 3, 10 February 2011, Pages 255–260
نویسندگان
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