Determination of recombinant protein in animal cell culture supernatant by near-infrared spectroscopy.
A method is described for the measurement of recombinant protein in animal cell culture supernatants with near-infrared (NIR) absorbance spectroscopy. Human antithrombin III is determined in CHO (Chinese hamster ovary)-T1 medium, both fresh and taken from CHO cell culture. NIR spectra are collected over the frequency range of 1000-2500 nm. A multivariate calibration model is generated by correlating spectral features with concentration values using partial least-squares regression. Separate test sets are used to evaluate the computed model. Protein measurement is established over the range 0.1-5 micrograms/mL with a standard error of prediction of less than 0.5 microgram/mL. This result underlines the analytical applicability of NIR spectroscopy for animal cell culture process monitoring.
Harthun
S
S
Technische Hochschule Darmstadt, Institut für Biochemie, Germany.
Matischak
K
K
Friedl
P
P
D000818:Animals; D000990:Antithrombin III / Q000032:analysis; D001709:Biotechnology; D016466:CHO Cells; D006224:Cricetinae; D017077:Culture Media, Conditioned; D006801:Humans; D016018:Least-Squares Analysis; D011994:Recombinant Proteins / Q000032:analysis*; D015203:Reproducibility of Results; D019265:Spectroscopy, Near-Infrared / Q000379:methods* / Q000706:statistics & numerical data
D017077:Culture Media, Conditioned; D011994:Recombinant Proteins; D000990:Antithrombin III