Fast quantification of recombinant protein inclusion bodies within intact cells by FT-IR spectroscopy.
The accomplishment of the quantification of the recombinant protein content of whole bacterial cells by FT-IR spectroscopy by application of chemometrics is shown. Recombinant Escherichia coli cells expressing an inclusion body forming fusion protein were dried on a 96-well silicon plate for the analysis in a high-throughput FT-IR spectrometer. Acquired spectra of additionally conventionally quantified samples were used to establish a multivariate calibration. The obtained method was tested by predicting inclusion body contents of samples not used for the multivariate model. Results from FT-IR spectra coincided well with the data of universalized electrophoresis analysis. Hence FT-IR spectroscopy could prove as a fast and simple alternative to conventional quantification methods.
Gross-Selbeck
Sven
S
Department of Biotechnology, University of Natural Resources, Applied Life Sciences, Vienna, Muthgasse 18, 1190 Vienna, Austria.
Margreiter
Gerd
G
Obinger
Christian
C
Bayer
Karl
K
D004926:Escherichia coli / Q000166:cytology / Q000378:metabolism; D002479:Inclusion Bodies / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism*; D015203:Reproducibility of Results; D017550:Spectroscopy, Fourier Transform Infrared / Q000379:methods*
D011994:Recombinant Proteins