Machine learning aided UV absorbance spectroscopy for microbial contamination in cell therapy products

Abstract We demonstrate the feasibility of machine-learning aided UV absorbance spectroscopy for in-process microbial contamination detection during cell therapy product (CTP) manufacturing. This method leverages a one-class support vector machine to analyse the absorbance spectra of cell cultures a...

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Main Authors: Shruthi Pandi Chelvam, Alice Jie Ying Ng, Jiayi Huang, Elizabeth Lee, Maciej Baranski, Derrick Yong, Rohan B. H. Williams, Stacy L. Springs, Rajeev J. Ram
Format: Article
Language:English
Published: Nature Portfolio 2025-03-01
Series:Scientific Reports
Online Access:https://doi.org/10.1038/s41598-024-83114-y
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author Shruthi Pandi Chelvam
Alice Jie Ying Ng
Jiayi Huang
Elizabeth Lee
Maciej Baranski
Derrick Yong
Rohan B. H. Williams
Stacy L. Springs
Rajeev J. Ram
author_facet Shruthi Pandi Chelvam
Alice Jie Ying Ng
Jiayi Huang
Elizabeth Lee
Maciej Baranski
Derrick Yong
Rohan B. H. Williams
Stacy L. Springs
Rajeev J. Ram
author_sort Shruthi Pandi Chelvam
collection DOAJ
description Abstract We demonstrate the feasibility of machine-learning aided UV absorbance spectroscopy for in-process microbial contamination detection during cell therapy product (CTP) manufacturing. This method leverages a one-class support vector machine to analyse the absorbance spectra of cell cultures and predict if a sample is sterile or contaminated. This label-free technique provides a rapid output (< 30 minutes) with minimal sample preparation and volume (< 1 mL). Spiking of 7 microbial organisms into mesenchymal stromal cells supernatant aliquots from 6 commercial donors showed that contamination events could be detected at low inoculums of 10 CFUs with mean true positive and negative rates of 92.7% and 77.7% respectively. The true negative rate further improved to 92% after excluding samples from a single donor with anomalously high nicotinic acid. In cells spiked with 10 CFUs of E. coli, contamination was detected at the 21-hour timepoint, demonstrating comparable sensitivity to compendial USP < 71 > test (~ 24 hours). We hypothesize that spectral differences between nicotinic acid and nicotinamide in the UV region are the underlying mechanisms for contamination detection. This approach can be deployed as a preliminary test during different CTP manufacturing stages, for real-time, continuous culture monitoring enabling early detection of microbial contamination, assuring safety of CTP.
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spelling doaj-art-0863d25fd03040bf883a13e650f67a872025-08-20T02:59:24ZengNature PortfolioScientific Reports2045-23222025-03-0115111410.1038/s41598-024-83114-yMachine learning aided UV absorbance spectroscopy for microbial contamination in cell therapy productsShruthi Pandi Chelvam0Alice Jie Ying Ng1Jiayi Huang2Elizabeth Lee3Maciej Baranski4Derrick Yong5Rohan B. H. Williams6Stacy L. Springs7Rajeev J. Ram8Critical Analytics for Manufacturing Personalized Medicine (CAMP), Singapore-MIT Alliance for Research and Technology CentreCritical Analytics for Manufacturing Personalized Medicine (CAMP), Singapore-MIT Alliance for Research and Technology CentreCritical Analytics for Manufacturing Personalized Medicine (CAMP), Singapore-MIT Alliance for Research and Technology CentreCritical Analytics for Manufacturing Personalized Medicine (CAMP), Singapore-MIT Alliance for Research and Technology CentreCritical Analytics for Manufacturing Personalized Medicine (CAMP), Singapore-MIT Alliance for Research and Technology CentreCritical Analytics for Manufacturing Personalized Medicine (CAMP), Singapore-MIT Alliance for Research and Technology CentreCritical Analytics for Manufacturing Personalized Medicine (CAMP), Singapore-MIT Alliance for Research and Technology CentreCritical Analytics for Manufacturing Personalized Medicine (CAMP), Singapore-MIT Alliance for Research and Technology CentreCritical Analytics for Manufacturing Personalized Medicine (CAMP), Singapore-MIT Alliance for Research and Technology CentreAbstract We demonstrate the feasibility of machine-learning aided UV absorbance spectroscopy for in-process microbial contamination detection during cell therapy product (CTP) manufacturing. This method leverages a one-class support vector machine to analyse the absorbance spectra of cell cultures and predict if a sample is sterile or contaminated. This label-free technique provides a rapid output (< 30 minutes) with minimal sample preparation and volume (< 1 mL). Spiking of 7 microbial organisms into mesenchymal stromal cells supernatant aliquots from 6 commercial donors showed that contamination events could be detected at low inoculums of 10 CFUs with mean true positive and negative rates of 92.7% and 77.7% respectively. The true negative rate further improved to 92% after excluding samples from a single donor with anomalously high nicotinic acid. In cells spiked with 10 CFUs of E. coli, contamination was detected at the 21-hour timepoint, demonstrating comparable sensitivity to compendial USP < 71 > test (~ 24 hours). We hypothesize that spectral differences between nicotinic acid and nicotinamide in the UV region are the underlying mechanisms for contamination detection. This approach can be deployed as a preliminary test during different CTP manufacturing stages, for real-time, continuous culture monitoring enabling early detection of microbial contamination, assuring safety of CTP.https://doi.org/10.1038/s41598-024-83114-y
spellingShingle Shruthi Pandi Chelvam
Alice Jie Ying Ng
Jiayi Huang
Elizabeth Lee
Maciej Baranski
Derrick Yong
Rohan B. H. Williams
Stacy L. Springs
Rajeev J. Ram
Machine learning aided UV absorbance spectroscopy for microbial contamination in cell therapy products
Scientific Reports
title Machine learning aided UV absorbance spectroscopy for microbial contamination in cell therapy products
title_full Machine learning aided UV absorbance spectroscopy for microbial contamination in cell therapy products
title_fullStr Machine learning aided UV absorbance spectroscopy for microbial contamination in cell therapy products
title_full_unstemmed Machine learning aided UV absorbance spectroscopy for microbial contamination in cell therapy products
title_short Machine learning aided UV absorbance spectroscopy for microbial contamination in cell therapy products
title_sort machine learning aided uv absorbance spectroscopy for microbial contamination in cell therapy products
url https://doi.org/10.1038/s41598-024-83114-y
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