Rapid Characterisation and Quality Control of Complex Cell Culture Media Solutions using Raman Spectroscopy and Chemometrics. B. Li, P.W. Ryan, B.H. Ray, K.J. Leister, N.M.S. Sirimuthu, and A.G. Ry Biotechnology and Bioengineering , 107(2), 290-301, (2010). DOI: 10.1002/bit.22813 . 1 RA P I D CH A R A C T E R I S A T I O N A N D QU A L I T Y C O N T R O L O F C O M P L E X C E L L C U L T U R E ME D I A SO L U T I O N S U S I N G RA M A N SP E C T R O S C O P Y A N D CH E M O M E T R I C S . Boyan Li, 1,2 Paul W. Ryan, 1,2 Bryan H. Ray, 1,2 Kirk J. Leister, 3 Narayana M. S. Sirimuthu, 1,2 and Alan G. Ryder. 1,2 * 1 Nanoscale Biophotonics Laboratory, School of Chemistry, National University of Ire Galway, Galway, Ireland. 2 Centre for Bioanalytical Sciences, School of Chemistry, National University of Irela Galway, Galway, Ireland. 3 Analytical Biochemistry Department, Bristol-Myers Squibb Company, East Syra NY, USA. * To whom all correspondence should be addressed. Tel: +353 91 49 2943 Fax: +353 91 49 4596 Email: alan.ryder@nuigalway.ie (A.G Running Title: Raman spectroscopic analysis of cell culture media. Abstract: The use of Raman spectroscopy coupled with chemometrics for the rapid identifica characterisation, and quality assessment of complex cell culture media components us industrial mammalian cell culture was investigated. Raman spectroscopy offers signifi advantages for the analysis of complex, aqueous based materials used in biotec because there is no need for sample preparation and water is a weak Raman scatterer demonstrate the efficacy of the method for the routine analysis of dilute aqueous solut five different chemically defined, commercial media components used in a Chinese Ha Ovary (CHO) cell manufacturing process for recombinant proteins. The chemometric processing of the Raman spectral data is the key factor in develo robust methods. Here we discuss the optimum methods for eliminating baselin background fluctuations and other instrumentation artefacts to generate reproducible data. Principal component analysis (PCA) and soft independent modelling of class ana (SIMCA) were then employed in the development of a robust routine for both identific and quality evaluation of the five different media components. These methods potential to be extremely useful in an industrial context for “in house” sample tracking and quality control. Keywords: Raman spectroscopy, chemometrics, cell culture, chemically defined eRDF, PCA, SIMCA. Final submitted version, not proof corrected.