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Antibodies 2017, 6(4), 24; https://doi.org/10.3390/antib6040024

Process Analytical Approach towards Quality Controlled Process Automation for the Downstream of Protein Mixtures by Inline Concentration Measurements Based on Ultraviolet/Visible Light (UV/VIS) Spectral Analysis

1
Institute for Separation and Process Technology, Clausthal University of Technology, Leibnizstraße 15, 38678 Clausthal-Zellerfeld, Germany
2
Institute for Process Control, Clausthal University of Technology, Arnold-Sommerfeld-Straße 1, 38678 Clausthal-Zellerfeld, Germany
*
Author to whom correspondence should be addressed.
Received: 31 October 2017 / Revised: 29 November 2017 / Accepted: 6 December 2017 / Published: 12 December 2017
(This article belongs to the Special Issue Monoclonal Antibodies)
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Abstract

Downstream of pharmaceutical proteins, such as monoclonal antibodies, is mainly done by chromatography, where concentration determination of coeluting components presents a major problem. Inline concentration measurements (ICM) by Ultraviolet/Visible light (UV/VIS)-spectral data analysis provide a label-free and noninvasive approach to significantly speed up the analysis and process time. Here, two different approaches are presented. For a test mixture of three proteins, a fast and easily calibrated method based on the non-negative least-squares algorithm is shown, which reduces the calibration effort compared to a partial least-squares approach. The accuracy of ICM for analytical separations of three proteins on an ion exchange column is over 99%, compared to less than 85% for classical peak area evaluation. The power of the partial least squares algorithm (PLS) is shown by measuring the concentrations of Immunoglobulin G (IgG) monomer and dimer under a worst-case scenario of completely overlapping peaks. Here, the faster SIMPLS algorithm is used in comparison to the nonlinear iterative partial least squares (NIPALS) algorithm. Both approaches provide concentrations as well as purities in real-time, enabling live-pooling decisions based on product quality. This is one important step towards advanced process automation of chromatographic processes. Analysis time is less than 100 ms and only one program is used for all the necessary communications and calculations. View Full-Text
Keywords: inline concentration measurements; UV/VIS spectral analysis; PAT for monoclonal antibodies; live pooling; peak deconvolution inline concentration measurements; UV/VIS spectral analysis; PAT for monoclonal antibodies; live pooling; peak deconvolution
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).
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Zobel-Roos, S.; Mouellef, M.; Siemers, C.; Strube, J. Process Analytical Approach towards Quality Controlled Process Automation for the Downstream of Protein Mixtures by Inline Concentration Measurements Based on Ultraviolet/Visible Light (UV/VIS) Spectral Analysis. Antibodies 2017, 6, 24.

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