title: Fourier transform assisted deconvolution of skewed peaks in complex multi-dimensional chromatograms.
abstract: Lower order peak moments of individual peaks in heavily fused peak clusters can be determined by fitting peak models to the experimental data. The success of such an approach depends on two main aspects: the generation of meaningful initial estimates on the number and position of the peaks, and the choice of a suitable peak model. For the detection of meaningful peaks in multi-dimensional chromatograms, a fast data scanning algorithm was combined with prior resolution enhancement through the reduction of column and system broadening effects with the help of two-dimensional fast Fourier transforms. To capture the shape of skewed peaks in multi-dimensional chromatograms a formalism for the accurate calculation of exponentially modified Gaussian peaks, one of the most popular models for skewed peaks, was extended for direct fitting of two-dimensional data. The method is demonstrated to successfully identify and deconvolute peaks hidden in strongly fused peak clusters. Incorporation of automatic analysis and reporting of the statistics of the fitted peak parameters and calculated properties allows to easily identify in which regions of the chromatograms additional resolution is required for robust quantification.
authors.affiliation: Department of Biotechnology, TU Delft, Julianalaan 67, 2628 BC, Delft, The Netherlands.
authors.forename: Alexander T
authors.initials: AT
authors.lastname: Hanke
authors.affiliation: Department of Biotechnology, TU Delft, Julianalaan 67, 2628 BC, Delft, The Netherlands.
authors.forename: Peter D E M
authors.initials: PD
authors.lastname: Verhaert
authors.affiliation: Department of Biotechnology, TU Delft, Julianalaan 67, 2628 BC, Delft, The Netherlands.
authors.forename: Luuk A M
authors.initials: LA
authors.lastname: van der Wielen
authors.affiliation: Synthon Biopharmaceuticals B.V., Microweg 22, 6503 GN, Nijmegen, The Netherlands.
authors.forename: Michel H M
authors.initials: MH
authors.lastname: Eppink
authors.affiliation: DSM Biotechnology Center, Alexander Fleminglaan 1, 2613 AX, Delft, The Netherlands.
authors.forename: Emile J A X
authors.initials: EJ
authors.lastname: van de Sandt
authors.affiliation: Department of Biotechnology, TU Delft, Julianalaan 67, 2628 BC, Delft, The Netherlands. Electronic address: m.ottens@tudelft.nl.
authors.forename: Marcel
authors.initials: M
authors.lastname: Ottens
mesh_terms: D000465:Algorithms; D000818:Animals; D016466:CHO Cells; D002850:Chromatography, Gel / Q000379:methods*; D003412:Cricetulus; D005583:Fourier Analysis; D007074:Immunoglobulin G / Q000032:analysis; D008962:Models, Theoretical
chemical_list: D007074:Immunoglobulin G
keywords: Comprehensive two-dimensional chromatography; Deconvolution; Exponentially modified Gaussian; Fourier transform; Non-linear curve fitting; Peak model