A rapid method for the differentiation of yeast cells grown under carbon and nitrogen-limited conditions by means of partial least squares discriminant analysis employing infrared micro-spectroscopic data of entire yeast cells

TALANTA

卷:99

Article

Kuligowski, Julia²  Quintas, Guillermo³  Herwig, Christoph⁴  Lendl, Bernhard¹ 

[1] Vienna Univ Technol, Inst Chem Technol & Analyt, A-1060 Vienna, Austria

[2] Univ Valencia, Dept Analyt Chem, E-46100 Burjassot, Spain

[3] Leitat Technol Ctr, Bio InVitro Div, E-08225 Terrassa, Spain

[4] Vienna Univ Technol, Inst Chem Engn, Res Area Biochem Engn, A-1060 Vienna, Austria

关键词: Infrared (IR) micro-spectroscopy;    Partial least squares-discriminant analysis (PLS-DA);    Permutation testing;    Double cross validation (2CV);    Pichia pastoris;    Yeast;   

DOI  :  10.1016/j.talanta.2012.06.036

来源: Elsevier

【 摘 要 】

This paper shows the ease of application and usefulness of mid-IR measurements for the investigation of orthogonal cell states on the example of the analysis of Pichia pastoris cells. A rapid method for the discrimination of entire yeast cells grown under carbon and nitrogen-limited conditions based on the direct acquisition of mid-IR spectra and partial least squares discriminant analysis (PLS-DA) is described. The obtained PLS-DA model was extensively validated employing two different validation strategies: (i) statistical validation employing a method based on permutation testing and (ii) external validation splitting the available data into two independent sub-sets. The Variable Importance in Projection scores of the PLS-DA model provided deeper insight into the differences between the two investigated states. Hence, we demonstrate the feasibility of a method which uses IR spectra from intact cells that may be employed in a second step as an in-line tool in process development and process control along Quality by Design principles. (C) 2012 Elsevier B.V. All rights reserved.

【 授权许可】

Free