Gross-Selbeck Sven, Margreiter Gerd, Obinger Christian, Bayer Karl
Department of Biotechnology, University of Natural Resources, Applied Life Sciences, Vienna, Muthgasse 18, 1190 Vienna, Austria.
Biotechnol Prog. 2007 May-Jun;23(3):762-6. doi: 10.1021/bp070022q. Epub 2007 May 11.
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.
展示了通过化学计量学应用傅里叶变换红外光谱法对全细菌细胞中重组蛋白含量进行定量的成果。将表达形成包涵体融合蛋白的重组大肠杆菌细胞干燥在96孔硅板上，以便在高通量傅里叶变换红外光谱仪中进行分析。使用另外常规定量样品的获取光谱来建立多元校准。通过预测未用于多元模型的样品的包涵体含量来测试所获得的方法。傅里叶变换红外光谱的结果与通用电泳分析的数据非常吻合。因此，傅里叶变换红外光谱法可以证明是一种快速简单的传统定量方法的替代方法。
