Mammalian Cell culture technology is the primary means of manufacturing biotherapeutics including monoclonal antibodies. There are a variety of biotherapeutics that are produced in small and large-scale processes. This can be for the purposes of development, clinical studies, or the production of marketed products.

Federal regulatory bodies are recommending the use of reliable Process Analytical Technology (PAT) to ensure the reliability of their processes. Process Analytical Technologies provide feedback while the process is running, allowing for real-time adjustments to ensure quality. This includes upstream cell culture manufacturing processes. This has driven companies operating in Current Good Manufacturing Practice (cGMP) environments to look into the integration of Raman as a PAT tool in order to comply with the recommendations and quality expectations.

Raman spectroscopy technology is an ideal tool for upstream PAT as it has been shown to facilitate the monitoring of several important CQAs within mammalian cell cultures. Therefore it provides an important means of monitoring and/or controlling bioprocesses. The following are some examples of CQAs of interest for Raman measurements with Tornado’s Raman spectroscopy-based analyzers:

• Glucose
• Lactate
• Viable Cell Count and Total Cell count (VCC and TCC)
• Protein titer
• Glutamine, Glutamate, and other amino acids
• Acetate
• Ammonium

Cell culture process development, optimization, and scaling are seminal steps in producing biopharmaceutical therapeutics, such as monoclonal antibodies. Continuous in-line, real-time measurements can facilitate process understanding, which allows process optimization. However, in-line, real-time measurements can be challenging to obtain with enough sensitivity and accuracy to be useful. The superior performance of Tornado’s Raman spectrometers can help overcome these limitations, thus making it an ideal solution for cell culture process development.

Learn how Tornado’s Raman Analyzers are compatible with Sartorius Ambr® and STR® platforms through seamless integration with RamanBioConnect™ Raman Probe.

The coupling of the ultrasound technique with Tornado’s Raman spectroscopy-based analyzer offers a unique capability – in-situ sample manipulation to obtain critical information about the solid and solution fractions in a cell reactor, i.e., the microorganisms and media/milieu.

By utilizing ultrasonic manipulation, particles of the solid fraction can be manipulated in a couple of ways. They can be sequestered by the standing wave in the focal plane of the immersion Raman probe or they can be cleared from the focal plane to enhance the spectrum of the solution phase. Trapping or clearing of the particles of interest allows for information to be gathered on phase individually.

The increased signal strength of the HTVS™ technology allows for enhanced sensitivity and selectivity in the measurement, thus, valuable information can be obtained, leading to better process understanding. The details of work done with ultrasonic manipulation can be found in this white paper: Coupled High-Throughput Raman and Ultrasonic Particle Manipulation for Increased Sensitivity and Selectivity in a Cell Reactor.

Raman spectroscopy has proven advantages for process monitoring and control of batch cultivations. However, there is a growing trend in the industry to move toward perfusion processes that allow greater efficiency and economy in the production of large molecules. Tornado’s Raman spectroscopy offers opportunities for advanced process monitoring and control of perfusion processes, bringing advantages similar to what it imparts for batch processes.

Tornado’s Raman spectroscopy-based analyzers with their revolutionary, patented spectrometer technology can be used as PAT tools for in-line CQA measurements in prolonged perfusion processes. This is especially true in the critical aspect of nutrient control, even if low and stable glucose concentrations are required. Read more about this in the following partner publication.

Chinese hamster ovary (CHO) cells are the preferred production platform for therapeutic monoclonal antibodies (mAb) due to their ability to produce antibodies that are similar in glycosylation to human serum antibodies.

Tornado’s Raman spectroscopy-based analyzers are exceptionally suited to monitor critical process parameters (CCP) through off-line, at-line, and in-line real-time monitoring of titer, total and viable cell counts/densities, glucose, lactate, osmolality, ammonium and antibody concentration in CHO cell culture for the production of monoclonal antibodies. Tornado’s HudsonTM Bioreactor Raman Immersion Probe provides an autoclavable solution for sterile in-line spectral acquisition, eliminating the risk of contamination from repeated sampling for off-line/at-line analysis.