title: Fast quantification of recombinant protein inclusion bodies within intact cells by FT-IR spectroscopy.
abstract: 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.
authors.affiliation: Department of Biotechnology, University of Natural Resources, Applied Life Sciences, Vienna, Muthgasse 18, 1190 Vienna, Austria.
authors.forename: Sven
authors.initials: S
authors.lastname: Gross-Selbeck
authors.forename: Gerd
authors.initials: G
authors.lastname: Margreiter
authors.forename: Christian
authors.initials: C
authors.lastname: Obinger
authors.forename: Karl
authors.initials: K
authors.lastname: Bayer
mesh_terms: 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*
chemical_list: D011994:Recombinant Proteins