Related Experiment Video
Updated: Jul 29, 2026
09:56
Real-time Monitoring of Reactions Performed Using Continuous-flow Processing: The Preparation of 3-Acetylcoumarin as an Example
Published on: November 18, 2015
10.2K
Flow Water Proton NMR: In-Line Process Analytical Technology for Continuous Biomanufacturing
Marc B Taraban1, Katharine T Briggs1, Peter Merkel1
1Department of Pharmaceutical Sciences, School of Pharmacy , University of Maryland , Baltimore , Maryland 21201 , United States.
Analytical Chemistry
|September 25, 2019
Summary
A new water proton NMR method (flow-wNMR) enables real-time, in-line monitoring of protein concentration and aggregate levels in continuous biomanufacturing. This noninvasive process analytical technology (PAT) addresses a key challenge for the biopharmaceutical industry.
Area of Science:
- Biopharmaceutical Manufacturing
- Process Analytical Technology (PAT)
- Nuclear Magnetic Resonance (NMR) Spectroscopy
Background:
- Continuous manufacturing is a key goal for the biopharmaceutical industry.
- Current process analytical technology (PAT) lacks noninvasive, real-time monitoring for critical quality attributes like protein concentration and aggregates.
- Water proton transverse relaxation rate (R2(1H2O)) has shown sensitivity to protein concentration and aggregates in static biopharmaceutical formulations.
Purpose of the Study:
- To investigate the feasibility of using water proton NMR under flow conditions (flow-wNMR) for real-time, in-line monitoring.
- To establish R2(1H2O) as a quantitative indicator for protein concentration and aggregate variations in flowing biopharmaceutical processes.
Main Methods:
- Development and application of a flow-wNMR system for monitoring biopharmaceutical formulations.
- Measurement of water proton transverse relaxation rate (R2(1H2O)) under dynamic flow conditions.
- Correlation of R2(1H2O) changes with variations in protein concentration and aggregate content.
Main Results:
- Flow-wNMR demonstrated sensitivity to small changes in protein concentration, below 1 mg/mL.
- The technique successfully detected variations in aggregate content below 1%.
- R2(1H2O) was confirmed as a reliable quantitative indicator in a dynamic flow system.
Conclusions:
- Flow-wNMR is a promising noninvasive process analytical technology (PAT) for continuous biomanufacturing.
- This method enables real-time, in-line monitoring of critical product quality attributes.
- Implementation of flow-wNMR can significantly advance the adoption of continuous bioprocessing.
Related Concept Videos
01:14Batch vs Continuous Culture
Fermentation is a foundational biotechnological process used to produce pharmaceuticals, biofuels, enzymes, and food additives. Among industrial strategies, batch and continuous fermentation are the two most widely applied. Although both rely on microbial conversion of substrates into desired products, they differ markedly in operation, productivity, and suitability for specific applications.Batch fermentation occurs in a closed system in which nutrient media and inoculum are added at the...
01:14Scale-Up Processes
The scale-up of microbial fermentation processes is essential in industrial biotechnology, allowing the transition from laboratory-scale experiments to commercial-scale production while aiming to maintain product yield and quality. This process requires meticulous adjustment of equipment design, process parameters, and contamination control strategies to accommodate increasing culture volumes.At the laboratory scale, cultures are typically maintained in 1 to 10-liter glass or autoclavable...
01:27Upstream Processing
Upstream processing represents a critical phase in biomanufacturing, wherein biological systems such as microorganisms, mammalian cells, or insect cells are cultivated to produce therapeutic proteins, vaccines, enzymes, or other biologically derived products. This phase encompasses all steps from the selection and genetic manipulation of the production organism to the cultivation of cells in bioreactors under tightly controlled environmental conditions.Host Selection and Genetic OptimizationThe...
01:27Production of Alcohol
Continuous fermentation is a key strategy in industrial ethanol production, particularly when efficiency, scalability, and high yields are essential. This approach allows for uninterrupted operation and optimized resource utilization. The primary feedstock, corn starch, undergoes enzymatic hydrolysis facilitated by α-amylase and glucoamylase. These enzymes break down the starch into fermentable sugars such as glucose, which are readily assimilated by fermentative microorganisms.Fermentation...
01:29Downstream Processing
Downstream processing begins once fermentation is complete and involves a series of steps to recover and purify products such as acids, vitamins, antibiotics, or proteins.Cell HarvestingFor example, for intracellular protein-based products, the first step is harvesting the cells. This is typically achieved using centrifugation or filtration to separate the cells from the liquid phase.Cell Disruption for Intracellular ProductsIf the target product is intracellular, the harvested cells must be...