A Modular Mechanistic In Silico Model for In Vitro Transcription Process Yield and Product Quality Prediction

Fuente: arXiv
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Autores principales: Wang, Keqi, Choy, Keilung, Reiser, Eli, Pei, Jinxiang, Zheng, Hua, Dasgupta, Aparajita, Cheng, Fuqiang, Dong, Guogang, Mulukutla, Bhanu Chandra, Mannheimer, Joshua, Huang, Carolyn, Farsani, Hooman, Xie, Wei
Formato: Preprint
Publicado: 2026
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author Wang, Keqi
Choy, Keilung
Reiser, Eli
Pei, Jinxiang
Zheng, Hua
Dasgupta, Aparajita
Cheng, Fuqiang
Dong, Guogang
Mulukutla, Bhanu Chandra
Mannheimer, Joshua
Huang, Carolyn
Farsani, Hooman
Xie, Wei
author_facet Wang, Keqi
Choy, Keilung
Reiser, Eli
Pei, Jinxiang
Zheng, Hua
Dasgupta, Aparajita
Cheng, Fuqiang
Dong, Guogang
Mulukutla, Bhanu Chandra
Mannheimer, Joshua
Huang, Carolyn
Farsani, Hooman
Xie, Wei
contents In vitro transcription (IVT) plays a critical role in the manufacture of mRNA vaccines and therapeutics. Optimizing mRNA yield and ensuring product quality, such as capping efficiency and integrity, are essential but mechanistically complex. This study presents a modular mechanistic model of the IVT process to advance scientific understanding and improve predictive capability. The IVT reaction network is decomposed into interconnected modules describing (1) initiation and capping, (2) elongation and truncation, (3) termination and read-through, (4) mRNA degradation, (5) magnesium pyrophosphate precipitation, and (6) enzymatic degradation of pyrophosphate. Guided by biochemical principles and experimental data, kinetic models were developed for each module, accounting for mass balances, molecular complexation, and enzyme activity, and were subsequently assembled to capture coupled IVT dynamics. Multivariate residual analysis and Shapley value-based sensitivity analysis, guided by domain knowledge, were applied to iteratively improve model fidelity. These machine learning-driven analytics enabled identification of key mechanisms, supported in silico experimentation, and facilitated root-cause analysis. Combined with Gaussian-process-based batch Bayesian optimization for efficient parameter estimation, this framework establishes a scalable hybrid (mechanistic + machine learning) modeling platform that integrates heterogeneous data, accelerates model calibration, and supports rational design and optimization of mRNA manufacturing processes.
format Preprint
id arxiv_https___arxiv_org_abs_2602_07257
institution arXiv
publishDate 2026
record_format arxiv
spellingShingle A Modular Mechanistic In Silico Model for In Vitro Transcription Process Yield and Product Quality Prediction
Wang, Keqi
Choy, Keilung
Reiser, Eli
Pei, Jinxiang
Zheng, Hua
Dasgupta, Aparajita
Cheng, Fuqiang
Dong, Guogang
Mulukutla, Bhanu Chandra
Mannheimer, Joshua
Huang, Carolyn
Farsani, Hooman
Xie, Wei
Molecular Networks
In vitro transcription (IVT) plays a critical role in the manufacture of mRNA vaccines and therapeutics. Optimizing mRNA yield and ensuring product quality, such as capping efficiency and integrity, are essential but mechanistically complex. This study presents a modular mechanistic model of the IVT process to advance scientific understanding and improve predictive capability. The IVT reaction network is decomposed into interconnected modules describing (1) initiation and capping, (2) elongation and truncation, (3) termination and read-through, (4) mRNA degradation, (5) magnesium pyrophosphate precipitation, and (6) enzymatic degradation of pyrophosphate. Guided by biochemical principles and experimental data, kinetic models were developed for each module, accounting for mass balances, molecular complexation, and enzyme activity, and were subsequently assembled to capture coupled IVT dynamics. Multivariate residual analysis and Shapley value-based sensitivity analysis, guided by domain knowledge, were applied to iteratively improve model fidelity. These machine learning-driven analytics enabled identification of key mechanisms, supported in silico experimentation, and facilitated root-cause analysis. Combined with Gaussian-process-based batch Bayesian optimization for efficient parameter estimation, this framework establishes a scalable hybrid (mechanistic + machine learning) modeling platform that integrates heterogeneous data, accelerates model calibration, and supports rational design and optimization of mRNA manufacturing processes.
title A Modular Mechanistic In Silico Model for In Vitro Transcription Process Yield and Product Quality Prediction
topic Molecular Networks
url https://arxiv.org/abs/2602.07257