title: Rapid high-throughput characterisation, classification and selection of recombinant mammalian cell line phenotypes using intact cell MALDI-ToF mass spectrometry fingerprinting and PLS-DA modelling.
abstract: Despite many advances in the generation of high producing recombinant mammalian cell lines over the last few decades, cell line selection and development is often slowed by the inability to predict a cell line's phenotypic characteristics (e.g. growth or recombinant protein productivity) at larger scale (large volume bioreactors) using data from early cell line construction at small culture scale. Here we describe the development of an intact cell MALDI-ToF mass spectrometry fingerprinting method for mammalian cells early in the cell line construction process whereby the resulting mass spectrometry data are used to predict the phenotype of mammalian cell lines at larger culture scale using a Partial Least Squares Discriminant Analysis (PLS-DA) model. Using MALDI-ToF mass spectrometry, a library of mass spectrometry fingerprints was generated for individual cell lines at the 96 deep well plate stage of cell line development. The growth and productivity of these cell lines were evaluated in a 10L bioreactor model of Lonza's large-scale (up to 20,000L) fed-batch cell culture processes. Using the mass spectrometry information at the 96 deep well plate stage and phenotype information at the 10L bioreactor scale a PLS-DA model was developed to predict the productivity of unknown cell lines at the 10L scale based upon their MALDI-ToF fingerprint at the 96 deep well plate scale. This approach provides the basis for the very early prediction of cell lines' performance in cGMP manufacturing-scale bioreactors and the foundation for methods and models for predicting other mammalian cell phenotypes from rapid, intact-cell mass spectrometry based measurements.
authors.affiliation: Centre for Molecular Processing and School of Bioscience, University of Kent, Canterbury CT2 7NJ, UK.
authors.forename: Jane F
authors.initials: JF
authors.lastname: Povey
authors.affiliation: School of Chemical Engineering & Advanced Materials, Newcastle University, Newcastle upon Tyne NE1 7RU, UK.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: O'Malley
authors.affiliation: Lonza Biologics plc, 228 Bath Road, Slough SL1 4DX, UK.
authors.forename: Tracy
authors.initials: T
authors.lastname: Root
authors.affiliation: School of Chemical Engineering & Advanced Materials, Newcastle University, Newcastle upon Tyne NE1 7RU, UK.
authors.forename: Elaine B
authors.initials: EB
authors.lastname: Martin
authors.affiliation: School of Chemical Engineering & Advanced Materials, Newcastle University, Newcastle upon Tyne NE1 7RU, UK.
authors.forename: Gary A
authors.initials: GA
authors.lastname: Montague
authors.affiliation: Lonza Biologics plc, 228 Bath Road, Slough SL1 4DX, UK.
authors.forename: Marc
authors.initials: M
authors.lastname: Feary
authors.affiliation: Centre for Molecular Processing and School of Bioscience, University of Kent, Canterbury CT2 7NJ, UK.
authors.forename: Carol
authors.initials: C
authors.lastname: Trim
authors.affiliation: Lonza Biologics plc, 228 Bath Road, Slough SL1 4DX, UK.
authors.forename: Dietmar A
authors.initials: DA
authors.lastname: Lang
authors.affiliation: Lonza Biologics plc, 228 Bath Road, Slough SL1 4DX, UK.
authors.forename: Richard
authors.initials: R
authors.lastname: Alldread
authors.affiliation: Lonza Biologics plc, 228 Bath Road, Slough SL1 4DX, UK.
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Racher
authors.affiliation: Centre for Molecular Processing and School of Bioscience, University of Kent, Canterbury CT2 7NJ, UK. Electronic address: c.m.smales@kent.ac.uk.
authors.forename: C Mark
authors.initials: CM
authors.lastname: Smales
mesh_terms: D000818:Animals; D061249:Batch Cell Culture Techniques / Q000379:methods; D019149:Bioreactors; D016466:CHO Cells / Q000145:classification*; D006224:Cricetinae; D003412:Cricetulus; D016172:DNA Fingerprinting / Q000379:methods*; D016018:Least-Squares Analysis; D008322:Mammals / Q000276:immunology*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
keywords: Cell line development; Cell line prediction; Chinese hamster ovary cells; PLS-DA modelling; Whole cell MALDI-ToF mass spectrometry