Cargando…
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
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 l...
Autores principales: | , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2017
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6698811/ https://www.ncbi.nlm.nih.gov/pubmed/31548539 http://dx.doi.org/10.3390/antib6040024 |
_version_ | 1783444619139219456 |
---|---|
author | Zobel-Roos, Steffen Mouellef, Mourad Siemers, Christian Strube, Jochen |
author_facet | Zobel-Roos, Steffen Mouellef, Mourad Siemers, Christian Strube, Jochen |
author_sort | Zobel-Roos, Steffen |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-6698811 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-66988112019-09-05 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 Zobel-Roos, Steffen Mouellef, Mourad Siemers, Christian Strube, Jochen Antibodies (Basel) Article 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. MDPI 2017-12-12 /pmc/articles/PMC6698811/ /pubmed/31548539 http://dx.doi.org/10.3390/antib6040024 Text en © 2017 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Zobel-Roos, Steffen Mouellef, Mourad Siemers, Christian Strube, Jochen 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 |
title | 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 |
title_full | 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 |
title_fullStr | 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 |
title_full_unstemmed | 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 |
title_short | 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 |
title_sort | 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 |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6698811/ https://www.ncbi.nlm.nih.gov/pubmed/31548539 http://dx.doi.org/10.3390/antib6040024 |
work_keys_str_mv | AT zobelroossteffen processanalyticalapproachtowardsqualitycontrolledprocessautomationforthedownstreamofproteinmixturesbyinlineconcentrationmeasurementsbasedonultravioletvisiblelightuvvisspectralanalysis AT mouellefmourad processanalyticalapproachtowardsqualitycontrolledprocessautomationforthedownstreamofproteinmixturesbyinlineconcentrationmeasurementsbasedonultravioletvisiblelightuvvisspectralanalysis AT siemerschristian processanalyticalapproachtowardsqualitycontrolledprocessautomationforthedownstreamofproteinmixturesbyinlineconcentrationmeasurementsbasedonultravioletvisiblelightuvvisspectralanalysis AT strubejochen processanalyticalapproachtowardsqualitycontrolledprocessautomationforthedownstreamofproteinmixturesbyinlineconcentrationmeasurementsbasedonultravioletvisiblelightuvvisspectralanalysis |