Raman laser intensity and sample clarification on biochemical monitoring over Zika-VLP

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Cavalcante, Paulo Eduardo da Silva ; Rabello, Julia Publio ; Leme, Jaci ; Dias, Vinicius Aragao Tejo ; Barrence, Fernanda Angela Correia ; Guardalini, Luis Giovani de Oliveira ; Bernardino, Thaissa Consoni ; Almeida, Sabrina ; Tonso, Aldo ; Jorge, Soraia Attie Calil ; Nunez, Eutimio Gustavo Fernandez

• Document type: Journal article
• Source: Biochemical and Biophysical Research Communications; v. 733, p. 11-pg., 2024-09-19.

Abstract

In the current biopharmaceutical scenario, constant bioprocess monitoring is crucial for the quality and integrity of final products. Thus, process analytical techniques, such as those based on Raman spectroscopy, have been used as multiparameter tracking methods in pharma bioprocesses, which can be combined with chemometric tools, like Partial Least Squares (PLS) and Artificial Neural Networks (ANN). In some cases, applying spectra pre-processing techniques before modeling can improve the accuracy of chemometric model fittings to observed values. One of the biological applications of these techniques could have as a target the virus-like particles (VLP), a vaccine production platform for viral diseases. A disease that has drawn attention in recent years is Zika, with large-scale production sometimes challenging without an appropriate monitoring approach. This work aimed to define global models for Zika VLP upstream production monitoring with Raman considering different laser intensities (200 mW and 495 mW), sample clarification (with or without cells), spectra pre-processing approaches, and PLS and ANN modeling techniques. Six experiments were performed in a benchtop bioreactor to collect the Raman spectral and biochemical datasets for modeling calibration. The best models generated presented a mean absolute error and mean relative error respectively of 3.46 x 10(5) cell/mL and 35 % for viable cell density (Xv); 4.1 % and 5 % for cell viability (CV); 0.245 g/L and 3 % for glucose (Glc); 0.006 g/L and 18 % for lactate (Lac); 0.115 g/L and 26 % for glutamine (Gln); 0.132 g/L and 18 % for glutamate (Glu); 0.0029 g/L and 3 % for ammonium (NH4+); and 0.0103 g/L and 2 % for potassium (K+). Sample without conditioning (with cells) improved the models' adequacy, except for Glutamine. ANN better predicted CV, Gln, Glu, and K+, while Xv, Glc, Lac, and NH4+ presented no statistical difference between the chemometric tools. For most of the assessed experimental parameters, there was no statistical need for spectra pre-filtering, for which the models based on the raw spectra were selected as the best ones. Laser intensity impacts quality model predictions in some parameters, Xv, Gln, and K+ had a better performance with 200 mW of intensity (for PLS, ANN, and ANN, respectively), for CV the 495 mW laser intensity was better (for PLS), and for the other biochemical variables, the use of 200 or 495 mW did not impact model fitting adequacy. (AU)

• FAPESP's process: 23/09463-5 - Laser wavelength and sample conditioning effects on biochemical monitoring of SARS-CoV-2 VLP production upstream stage by Raman spectroscopy
• Grantee: Júlia Públio Rabello
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• FAPESP's process: 22/02713-3 - Establishment of scalable bioprocesses for producing virus-like particles
• Grantee: Eutimio Gustavo Fernández Núñez
• Support Opportunities: Research Grants - Initial Project