Forum for Science, Industry and Business

Sponsored by:     3M 
Search our Site:

 

Microfluidic molecular exchanger helps control therapeutic cell manufacturing

15.10.2018

Researchers have demonstrated an integrated technique for monitoring specific biomolecules - such as growth factors - that could indicate the health of living cell cultures produced for the burgeoning field of cell-based therapeutics.

Using microfluidic technology to advance the preparation of samples from the chemically complex bioreactor environment, the researchers have harnessed electrospray ionization mass spectrometry (ESI-MS) to provide online monitoring that they believe will provide for therapeutic cell production the kind of precision quality control that has revolutionized other manufacturing processes.


Close-up image shows the Dynamic Mass Spectrometry Probe developed to monitor the health of living cell cultures.

Credit: Rob Felt, Georgia Tech

"The way that the production of cell therapeutics is done today is very much an art," said Andrei Fedorov, Woodruff Professor in the George W. Woodruff School of Mechanical Engineering at the Georgia Institute of Technology.

"Process control must evolve very quickly to support the therapeutic applications that are emerging from bench science today. We think this technology will help us reach the goal of making these exciting cell-based therapies widely available."

By measuring very low concentrations of specific compounds secreted or excreted by cells, the technique could also help identify which biomolecules - of widely varying sizes - should be monitored to guide the control of cell health. Ultimately, the researchers hope to integrate their label-free monitoring directly into high-volume bioreactors that will produce cells in quantities large enough to make the new therapies available at a reasonable cost and consistent quality.

Development of the Dynamic Mass Spectrometry Probe (DMSP) was supported by the National Science Foundation (NSF) Engineering Research Center for Cell Manufacturing Technologies (CMaT), which is headquartered at Georgia Tech. The work was reported September 10 in the journal Biotechnology and Bioengineering.

Traditional ESI-MS techniques have revolutionized analytical chemistry by allowing precise identification of complex biological compounds. Because of complex sample preparation requirements, existing approaches to ESI-MS require too much time to be useful for continuous monitoring of cell growth in bioreactors, where maintaining narrow parameters for specific indicators of cellular health is critical. Biological samples also contain salts, which must be removed before introduction into the ESI-MS system.

To accelerate the analytical process, Fedorov and a team that included graduate research assistant Mason Chilmonczyk and research engineer Peter Kottke used microfluidic technology to help separate compounds of interest from the salts. Salt removal uses a monolithic device in which a size-selective membrane with nanoscale pores is placed between two fluid flows, one the chemically complex sample drawn from the bioreactors and the other salt-free water with conditioning compounds.

The smaller salt molecules readily diffuse out of the sampled bioreactor flow through the nanopores, while the larger biomolecules mostly remain for the subsequent ESI-MS analysis. Meanwhile, chemical additives are at the same time introduced into the sample mixture through the same membrane nanopores to enhance ionization of the target biomolecules in the sampled mixture for improved ESI-MS analysis.

"We have used advanced microfabrication techniques to create a microfluidic device that will be able to treat samples in less than a minute," said Chilmonczyk. "Traditional sample preparation can require hours to days."

The process can currently remove as much as 99 percent of the salt, while retaining 80 percent of the biomolecules. Introduction of the conditioning chemicals allows the molecules to accept a greater charge, improving the capability of the mass spectrometer to detect low concentration biomolecules, and to measure large molecules.

"We can detect really high molecular weight molecules that the mass spectrometer normally wouldn't be able to detect," Fedorov said. "The size difference in the molecules of interest can be dramatic, so the improvement in the limit of detection across a broad range of analyte molecular weights will allow this technique to be more useful in cell manufacturing."

Because they use state of the art microfabrication techniques, the DMSP devices can be mass produced, allowing sampling to be scaled up to include multiple bioreactors at low cost. The small size of the device channels - which are just five microns tall - allows the system to produce results with samples as small as 20 nanoliters - with the potential for reducing that to as little as a single nanoliter.

"We need to monitor small concentrations of large biomolecules in this messy environment in a production line in such a way that we can check at any point how the cells are doing," Fedorov said. "This system could continuously monitor whether certain molecules are excreted or secreted at a reduced or increased rate. By correlating these measurements with cell health and potency, we could improve the manufacturing process."

Before the analytical techniques can be applied to quality control, the researchers must first identify biomolecules that indicate health of the growing cells. By sampling the bioreactor content locally in the immediate vicinity of cells and allowing identification of very small quantities of biochemicals, the DMSP technology can help researchers identify changes in molecular concentrations - which range from pico-molar to micro-molar - that may indicate the state of cells in the bioreactors. This would prompt adjustment of conditions in a bioreactor just in time to return to the state of healthy cell growth.

"In this situation, we often can't see the trees for the forest," said Fedorov. "There is a lot of material available, but we are looking for just a handful of individual trees that indicate the health of the cells. Because the forest is overgrown, the few selected trees we need to examine are hard to find. This is a grand challenge technologically."

###

The research team also included Research Scientist Hazel Stevens and Professor Robert Guldberg, who is now at the University of Oregon.

Media Contact

John Toon
jtoon@gatech.edu
404-894-6986

 @GeorgiaTech

http://www.gatech.edu 

John Toon | EurekAlert!
Further information:
http://dx.doi.org/10.1002/bit.26832

More articles from Life Sciences:

nachricht Self-organizing molecules: Nanorings with two sides
24.07.2019 | Universität Duisburg-Essen

nachricht Genome research shows that the body controls the integrity of heritable genomes
24.07.2019 | Universität zu Köln

All articles from Life Sciences >>>

The most recent press releases about innovation >>>

Die letzten 5 Focus-News des innovations-reports im Überblick:

Im Focus: MOF@SAW: Nanoquakes and molecular sponges for weighing and separating tiny masses

Augsburg chemists and physicists report how they have succeeded in the extremely difficult separation of hydrogen and deuterium in a gas mixture.

Thanks to the Surface Acoustic Wave (SAW) technology developed here and already widely used, the University of Augsburg is internationally recognized as the...

Im Focus: Better thermal conductivity by adjusting the arrangement of atoms

Adjusting the thermal conductivity of materials is one of the challenges nanoscience is currently facing. Together with colleagues from the Netherlands and Spain, researchers from the University of Basel have shown that the atomic vibrations that determine heat generation in nanowires can be controlled through the arrangement of atoms alone. The scientists will publish the results shortly in the journal Nano Letters.

In the electronics and computer industry, components are becoming ever smaller and more powerful. However, there are problems with the heat generation. It is...

Im Focus: First-ever visualizations of electrical gating effects on electronic structure

Scientists have visualised the electronic structure in a microelectronic device for the first time, opening up opportunities for finely-tuned high performance electronic devices.

Physicists from the University of Warwick and the University of Washington have developed a technique to measure the energy and momentum of electrons in...

Im Focus: Megakaryocytes act as „bouncers“ restraining cell migration in the bone marrow

Scientists at the University Würzburg and University Hospital of Würzburg found that megakaryocytes act as “bouncers” and thus modulate bone marrow niche properties and cell migration dynamics. The study was published in July in the Journal “Haematologica”.

Hematopoiesis is the process of forming blood cells, which occurs predominantly in the bone marrow. The bone marrow produces all types of blood cells: red...

Im Focus: Artificial neural network resolves puzzles from condensed matter physics: Which is the perfect quantum theory?

For some phenomena in quantum many-body physics several competing theories exist. But which of them describes a quantum phenomenon best? A team of researchers from the Technical University of Munich (TUM) and Harvard University in the United States has now successfully deployed artificial neural networks for image analysis of quantum systems.

Is that a dog or a cat? Such a classification is a prime example of machine learning: artificial neural networks can be trained to analyze images by looking...

All Focus news of the innovation-report >>>

Anzeige

Anzeige

VideoLinks
Industry & Economy
Event News

2nd International Conference on UV LED Technologies & Applications – ICULTA 2020 | Call for Abstracts

24.06.2019 | Event News

SEMANTiCS 2019 brings together industry leaders and data scientists in Karlsruhe

29.04.2019 | Event News

Revered mathematicians and computer scientists converge with 200 young researchers in Heidelberg!

17.04.2019 | Event News

 
Latest News

Hidden dynamics detected in neuronal networks

23.07.2019 | Life Sciences

Towards a light driven molecular assembler

23.07.2019 | Life Sciences

A torque on conventional magnetic wisdom

23.07.2019 | Physics and Astronomy

VideoLinks
Science & Research
Overview of more VideoLinks >>>