Light Induced Fluorescence Sensing For Biotherapeutics Process Control Applications
Real-time light-induced fluorescence (LIF) has and continues to be an emerging process analytical technology (PAT) for biopharmaceutical GMP process monitoring and control. Its application span biotherapeutic modalities, intermediates, and other auto-fluorescent analytes of interest. LIF’s attraction is attributed to its unique superior sensitivity, low concentration (low dose) detection and rapid response capabilities compared with traditional process analysis approaches such as Raman, near infrared (NIR), and ultraviolet-visible (UV-Vis) spectroscopies. It can also often afford selective detection without a multivariate or chemometric model as a fluorescent active pharmaceutical ingredient (API) is often contained within a negligible emissive background comprised of non-emissive intermediates and excipients in in-process materials and final dosage formulations, respectively. Moreover, real-time LIF equipped with a suitable in situ probe is also applicable to liquids, solids, semi-solids, and turbid solutions.
Custom Sensor and Technology’s established photometric LIF sensor has been applied to routine process monitoring and control GMP applications. Real-time LIF has been used to monitor protein biotherapeutics, biogenic fluorophore (NADH, pyridoxin, tryptophan/tyrosine, FAD, and FMN) 1-3, protein folding, binding and lyophilization.4, 5 It has also found utility to detect real-time fouling in downstream biotherapeutic purification resins and assess water quality.6-9 Numerous applications are probable owing to the plethora of biopharmaceutical or biological relevant fluorophores.10-12
Our LIF sensor product solution can be either supplied with a front surface in-line probe or a single-use flow cell (Figure 1). It can be configured for up to two excitation-emission combinations and comes equipped with suitable solid or liquid performance standards. A photometric configuration enables the full power of LIF by leverages industrial leading LED technology for incident excitation intensity modulation to mitigate photodegradation. This is combined with suitable emission filtering and integrated detection to enable signal response optimization. Consequently, full-spectrum LIF are often overengineered for real-time applications as fluorescence is a broadband spectroscopy well suited for robust solid-state filter-based photometry. Moreover, full-spectrum LIF instrumentation are insufficient as they are often unable to modulate measurement parameters to optimize signal response, to thereby meet application criteria. A plethora of biologically relevant and biotherapeutic APIs exhibit native fluorescence where our LIF sensor has and continues to find utility.10-12
Summary of LIF applications:
- Biotherapeutic or biogenic fluorophore monitoring and quantitation
- Protein folding and binding
- Protein lyophilization
- Resin fouling detection
- In-process materials, drug substance, drug product liquids and turbid solutions
References
- Kaiser, C.; Pototzki, T.; Ellert, A.; Luttmann, R., Applications of PAT-Process Analytical Technology in Recombinant Protein Processes with Escherichia coli. Engineering in Life Sciences 2008, 8 (2), 132-138.
- Olsson, L.; Schulze, U.; Nielsen, J., On-line bioprocess monitoring - an academic discipline or an industrial tool? TrAC Trends in Analytical Chemistry 1998, 17 (2), 88-95.
- Rathore, A. S.; Bhambure, R.; Ghare, V., Process analytical technology (PAT) for biopharmaceutical products. Anal Bioanal Chem 2010.
- Hartono, D.; et al., Decorating Liquid Crystal Surfaces with Proteins for Real-Time Detection of Specific Protein-Protein Binding. Advanced Functional Materials 2009, NA.
- Ramachander, R.; Jiang, Y.; Li, C.; Eris, T.; Young, M.; Dimitrova, M.; Narhi, L., Solid state fluorescence of lyophilized proteins. Analytical Biochemistry 2008, 376 (2), 173-182.
- Ghervase, L.; Carstea, G.; Savastru, P. D., Laser Induced Fluoresence in Water Quality Assessment. Romanian Reports in Physics 2010, 62, 652–659.
- Research_Water Monitoring organic matter in drinking water systems using fluorescence spectroscopy; 2010.
- Sharikova, A. V.; Killinger, D. K., UV-Laser and LED Fluorescence Detection of Trace Organic Compounds in Drinking Water and Distilled Spirits. InTech: Croatia, 2010.
- 9Pollard, P. C., Fluorescence instrument for in situ monitoring of viral abundance in water, wastewater and recycled water. Journal of Virological Methods 2012, 181 (1), 97-102.
- Al-Saleh, W. M. Laser and Light induced fluorescence of Some Biofluorophores at Different pH. King Saud University, Riyadh-Saudi Arabia, 2008.
- Albert-Garcia, J. R.; Antón-Fos, G. M.; Duart, M. J.; Lahuerta Zamora, L.; Martínez Calatayud, J., Theoretical prediction of the native fluorescence of pharmaceuticals. Talanta 2009, 79 (2), 412-418.
- Paudel, A.; Raijada, D.; Rantanen, J., Raman spectroscopy in pharmaceutical product design. Advanced Drug Delivery Reviews 2015, 89, 3-20.