Selective high throughput protein quantification based on UV absorption spectra.

摘要: The application of high throughput experimentation (HTE) in protein purification process development has created an analytical bottleneck. Recently, a new label-free and non-invasive methodology for analyzing multicomponent mixtures by means spectral measurements was presented. Analytics based on the shown to increase selective quantification significantly, however this only demonstrated one particular combination. In work, possibilities limitations method are investigated further. Principal component analysis (PCA) performed broad range absorption spectra investigate their common characteristics differences. PCA used both cluster define measure similarity. For binary combinations, calibration precision decrease exponentially with defined similarity factor. Knowledge correlation can be determine priori whether will successful or not. Calibration robustness applying analytics liquid chromatography HTE mode. Further it shown, that difference 0.6% sufficient sucessfully preform two IgG1 monoclonals.