Monitoring small-scale bioreactor studies for media development using polarized total synchronous fluorescence spectroscopy (pTSFS) and synchronous light scattering (SyLS).
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| Title: | Monitoring small-scale bioreactor studies for media development using polarized total synchronous fluorescence spectroscopy (pTSFS) and synchronous light scattering (SyLS). |
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| Authors: | Boateng, Bernard O.1 (AUTHOR), Ryder, Alan G.1 (AUTHOR) alan.ryder@universityofgalway.ie |
| Source: | Journal of Biotechnology. Nov2024, Vol. 395, p205-215. 11p. |
| Subjects: | Biotechnological process monitoring, Light scattering, Support vector machines, Principal components analysis, Vector analysis |
| Abstract: | Biopharmaceutical process development often involves the use of small-scale bioreactors (SSBR) for optimizing media formulations and process conditions during scale up to commercial scale production. Two key process parameters (CPP) used in SSBR studies are protein titre and viable cell density (VCD). Here, we explore the efficacy of parallel polarized total synchronous fluorescence spectroscopy (TSFS ||) and Synchronous Light Scattering (SyLS ||) to qualitatively monitor these CPPs and quantitatively predict titre and VCD for a large-scale cell culture media optimization SSBR study. The study involved 71 different media formulations (50+ components each), and the bioprocess was run for 13 days or more. Samples were extracted at set times (Day 0, 3, 9, and 13) and clarified by centrifugation. TSFS || spectra showed significant emission changes along with increased light scatter over the course of the bioprocess. SyLS || measurements strongly correlated with particle size data obtained from Dynamic Light Scattering but did not correlate well with VCD probably because of the centrifugation-based sample preparation. Statistical and principal component analysis (PCA) of the pTSFS data showed that spectral variation was greater between media formulations than due to the evolving bioprocess. This prevented the development of accurate global prediction models for media performance (e.g., predicting product titre at day 9 from media spectra measured at day 0). However, classification methods were successfully used to select media subsets with better quantitative prediction accuracy based on spectral similarities. A practical binary (high/low performance) classification model based on Support Vector Machines was generated for media formulation screening. Combining emission and scatter measurements with multivariate data analysis provides a more holistic, multi-attribute bioprocess monitoring method that minimizes the need to use different offline analytical methods. This methodology can be used to monitor process trajectories and deviations, and ultimately be used to predict bioprocess CPPs when implemented on production scale processes where there is much less compositional variation in the media. We believe this SSBR-pTSFS/SyLS approach will provide a valuable resource to develop the design/parameter space for in-process monitoring at production scale from early-stage process/media development studies. • First use of polarised emission (TSFS ‖) for analysis of small scale bioreactors (SSBR). • First use of polarised synchronous light scattering (SyLS ‖) for analysis of SSBR. • Combination of TSFS ‖ and SyLS ‖ used to monitor bioprocesses. • Support vector machine analysis of polarised emission predicts media performance. [ABSTRACT FROM AUTHOR] |
| Copyright of Journal of Biotechnology is the property of Elsevier B.V. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.) | |
| Database: | Engineering Source |
| FullText | Text: Availability: 0 |
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| Header | DbId: egs DbLabel: Engineering Source An: 180531188 AccessLevel: 6 PubType: Academic Journal PubTypeId: academicJournal PreciseRelevancyScore: 0 |
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| Items | – Name: Title Label: Title Group: Ti Data: Monitoring small-scale bioreactor studies for media development using polarized total synchronous fluorescence spectroscopy (pTSFS) and synchronous light scattering (SyLS). – Name: Author Label: Authors Group: Au Data: <searchLink fieldCode="AR" term="%22Boateng%2C+Bernard+O%2E%22">Boateng, Bernard O.</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Ryder%2C+Alan+G%2E%22">Ryder, Alan G.</searchLink><relatesTo>1</relatesTo> (AUTHOR)<i> alan.ryder@universityofgalway.ie</i> – Name: TitleSource Label: Source Group: Src Data: <searchLink fieldCode="JN" term="%22Journal+of+Biotechnology%22">Journal of Biotechnology</searchLink>. Nov2024, Vol. 395, p205-215. 11p. – Name: Subject Label: Subjects Group: Su Data: <searchLink fieldCode="DE" term="%22Biotechnological+process+monitoring%22">Biotechnological process monitoring</searchLink><br /><searchLink fieldCode="DE" term="%22Light+scattering%22">Light scattering</searchLink><br /><searchLink fieldCode="DE" term="%22Support+vector+machines%22">Support vector machines</searchLink><br /><searchLink fieldCode="DE" term="%22Principal+components+analysis%22">Principal components analysis</searchLink><br /><searchLink fieldCode="DE" term="%22Vector+analysis%22">Vector analysis</searchLink> – Name: Abstract Label: Abstract Group: Ab Data: Biopharmaceutical process development often involves the use of small-scale bioreactors (SSBR) for optimizing media formulations and process conditions during scale up to commercial scale production. Two key process parameters (CPP) used in SSBR studies are protein titre and viable cell density (VCD). Here, we explore the efficacy of parallel polarized total synchronous fluorescence spectroscopy (TSFS ||) and Synchronous Light Scattering (SyLS ||) to qualitatively monitor these CPPs and quantitatively predict titre and VCD for a large-scale cell culture media optimization SSBR study. The study involved 71 different media formulations (50+ components each), and the bioprocess was run for 13 days or more. Samples were extracted at set times (Day 0, 3, 9, and 13) and clarified by centrifugation. TSFS || spectra showed significant emission changes along with increased light scatter over the course of the bioprocess. SyLS || measurements strongly correlated with particle size data obtained from Dynamic Light Scattering but did not correlate well with VCD probably because of the centrifugation-based sample preparation. Statistical and principal component analysis (PCA) of the pTSFS data showed that spectral variation was greater between media formulations than due to the evolving bioprocess. This prevented the development of accurate global prediction models for media performance (e.g., predicting product titre at day 9 from media spectra measured at day 0). However, classification methods were successfully used to select media subsets with better quantitative prediction accuracy based on spectral similarities. A practical binary (high/low performance) classification model based on Support Vector Machines was generated for media formulation screening. Combining emission and scatter measurements with multivariate data analysis provides a more holistic, multi-attribute bioprocess monitoring method that minimizes the need to use different offline analytical methods. This methodology can be used to monitor process trajectories and deviations, and ultimately be used to predict bioprocess CPPs when implemented on production scale processes where there is much less compositional variation in the media. We believe this SSBR-pTSFS/SyLS approach will provide a valuable resource to develop the design/parameter space for in-process monitoring at production scale from early-stage process/media development studies. • First use of polarised emission (TSFS ‖) for analysis of small scale bioreactors (SSBR). • First use of polarised synchronous light scattering (SyLS ‖) for analysis of SSBR. • Combination of TSFS ‖ and SyLS ‖ used to monitor bioprocesses. • Support vector machine analysis of polarised emission predicts media performance. [ABSTRACT FROM AUTHOR] – Name: AbstractSuppliedCopyright Label: Group: Ab Data: <i>Copyright of Journal of Biotechnology is the property of Elsevier B.V. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract.</i> (Copyright applies to all Abstracts.) |
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| RecordInfo | BibRecord: BibEntity: Identifiers: – Type: doi Value: 10.1016/j.jbiotec.2024.10.002 Languages: – Code: eng Text: English PhysicalDescription: Pagination: PageCount: 11 StartPage: 205 Subjects: – SubjectFull: Biotechnological process monitoring Type: general – SubjectFull: Light scattering Type: general – SubjectFull: Support vector machines Type: general – SubjectFull: Principal components analysis Type: general – SubjectFull: Vector analysis Type: general Titles: – TitleFull: Monitoring small-scale bioreactor studies for media development using polarized total synchronous fluorescence spectroscopy (pTSFS) and synchronous light scattering (SyLS). Type: main BibRelationships: HasContributorRelationships: – PersonEntity: Name: NameFull: Boateng, Bernard O. – PersonEntity: Name: NameFull: Ryder, Alan G. IsPartOfRelationships: – BibEntity: Dates: – D: 20 M: 11 Text: Nov2024 Type: published Y: 2024 Identifiers: – Type: issn-print Value: 01681656 Numbering: – Type: volume Value: 395 Titles: – TitleFull: Journal of Biotechnology Type: main |
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