Abstract

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 IgG(4) 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.
Original languageUnknown
Pages (from-to)255-260
JournalJournal of Biotechnology
Volume151
Issue number3
DOIs
Publication statusPublished - 1 Jan 2011

Cite this

@article{3aa9fa3826cf4536b2bfaf90e94e72b5,
title = "High-throughput analysis of animal cell cultures using two-dimensional fluorometry",
abstract = "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 IgG(4) 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.",
keywords = "ANTIBODY-PRODUCTION, FLUORESCENCE SPECTROSCOPY, BIOPROCESS, BIOREACTOR",
author = "Oliveira, {Rui Manuel Freitas} and Carrondo, {Manuel Jos{\'e} Teixeira} and Alves, {Paula Maria} and Teixeira, {Ana Margarida}",
year = "2011",
month = "1",
day = "1",
doi = "10.1016/j.jbiotec.2010.11.015",
language = "Unknown",
volume = "151",
pages = "255--260",
journal = "Journal of Biotechnology",
issn = "0168-1656",
publisher = "Elsevier Science B.V., Inc",
number = "3",

}

TY - JOUR

T1 - High-throughput analysis of animal cell cultures using two-dimensional fluorometry

AU - Oliveira, Rui Manuel Freitas

AU - Carrondo, Manuel José Teixeira

AU - Alves, Paula Maria

AU - Teixeira, Ana Margarida

PY - 2011/1/1

Y1 - 2011/1/1

N2 - 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 IgG(4) 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.

AB - 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 IgG(4) 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.

KW - ANTIBODY-PRODUCTION

KW - FLUORESCENCE SPECTROSCOPY

KW - BIOPROCESS

KW - BIOREACTOR

U2 - 10.1016/j.jbiotec.2010.11.015

DO - 10.1016/j.jbiotec.2010.11.015

M3 - Article

VL - 151

SP - 255

EP - 260

JO - Journal of Biotechnology

JF - Journal of Biotechnology

SN - 0168-1656

IS - 3

ER -