Biochemical monitoring throughout all stages of rabies virus-like particles production by Raman spectroscopy using global models

Guardalini, Luis Giovani Oliveira; Cavalcante, Paulo Eduardo da Silva; Leme, Jaci; Mello, Renata Gois de; Bernardino, Thaissa Consoni; Astray, Renato Mancini; Barbosa, Eduardo Moreno; Silveira, Suellen Regina da; Ho, Paulo Lee; Tonso, Aldo; Jorge, Soraia Attie Calil; Núñez, Eutímio Gustavo Fernández

doi:10.1016/j.jbiotec.2022.12.009

Cited by 11 publications

(7 citation statements)

References 37 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This selection was performed using several spectral filtering techniques (as described in Table 2 and in previous studies.) by analysis of variance (ANOVA) and Tukey's significant difference test (HSD) 39 . The chosen comparative parameter was the MRE.…”

Section: Methodsmentioning

confidence: 99%

“…In another research analyzing global models in the biochemical monitoring of rabies VLP production stages using Raman spectroscopy in 3 L bioreactor assays, similar RMSE cv values were obtained for the same spectral ranges studied in the present work, particularly for the in line monitored sample database. 39 In Tables 3 and 4 can be observed the values of MAE ± SD AE were calculated for PLS. The absolute errors resulting from samples in the spectral range of 400-1850 cm −1 were lower than those obtained in the entire spectral range, with the in line sample database showing the lowest errors for all parameters, except for glutamate, which produced identical results for all databases in both spectral ranges.…”

Section: Pls Modelingmentioning

confidence: 99%

See 1 more Smart Citation

Monitoring by Raman spectroscopy of rabies virus‐like particles production since the initial development stages

Guardalini,

Rangel,

Leme

et al. 2023

J of Chemical Tech & Biotech

Self Cite

View full text Add to dashboard Cite

BACKGROUNDThis study aimed to establish chemometric models using Raman spectroscopy data for biochemical monitoring of rabies Virus‐Like Particles (VLP) production based on baculovirus/insect cell system. The models were developed using fresh and stored samples from the initial development stages (Schott culture flasks). The following modeling techniques were assessed: partial least squares (PLS) and artificial neural networks (ANN). The effects of spectral filtering approaches, spectral ranges (400–1850 cm−1; 100–3425 cm−1), and sample cryopreservation were also considered. The applicability of the models was evaluated using experimental data from assays carried out in a benchtop bioreactor.RESULTSThe results showed that the prediction capacity of the chemometric models was negatively impacted when samples from rabies VLP production were cryopreserved. Further studies are needed to confirm the maximum storage time for samples (< 4 months) without a significant difference in model predictions compared to those from an in line database. The dilution of the sample should be kept constant throughout the rabies VLP development stages. A nonlinear correlation was observed between dilution and the predicted values of biochemical parameters from Raman spectral data. The choice of spectral filtering has a major impact on the prediction accuracy of chemometric models.CONCLUSIONThe optimal filtering approach should be individually optimized for each biochemical parameter. The ANN models were significantly more suitable for biochemical monitoring than the PLS approach. The 400–1850 cm−1 Raman shift range is recommended for biochemical monitoring of rabies VLP using a baculovirus/insect cell platform when samples are cell‐free. © 2023 Society of Chemical Industry (SCI).

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Pls Modelingmentioning

confidence: 99%

Monitoring by Raman spectroscopy of rabies virus‐like particles production since the initial development stages

Guardalini,

Rangel,

Leme

et al. 2023

J of Chemical Tech & Biotech

Self Cite

View full text Add to dashboard Cite

show abstract

“…Data were diversified using different lysate batches and conditioning of the lysates by dilution, salt addition, or spiking with purified VLP or HCP solutions. Similar strategies for data diversification were employed in the literature by spiking purified monoclonal antibodies with mock solutions to mimic complex feedstocks (Großhans et al, 2019), spiking with nutrients during cell culture (Santos et al, 2018), spiking with cells during inoculum maturation (Guardalini et al, 2023b) or mixing available samples (Wang et al, 2023). In general, diversification strategies were reported to improve model robustness and interpretability (Santos et al, 2018) or enlarge the experimental data set (Wang et al, 2023).…”

Section: Effects Of Data Diversificationmentioning

confidence: 98%

Raman-based PAT for VLP precipitation: systematic data diversification and preprocessing pipeline identification

Dietrich,

Schiemer,

Kurmann

et al. 2024

Front. Bioeng. Biotechnol.

View full text Add to dashboard Cite

Virus-like particles (VLPs) are a promising class of biopharmaceuticals for vaccines and targeted delivery. Starting from clarified lysate, VLPs are typically captured by selective precipitation. While VLP precipitation is induced by step-wise or continuous precipitant addition, current monitoring approaches do not support the direct product quantification, and analytical methods usually require various, time-consuming processing and sample preparation steps. Here, the application of Raman spectroscopy combined with chemometric methods may allow the simultaneous quantification of the precipitated VLPs and precipitant owing to its demonstrated advantages in analyzing crude, complex mixtures. In this study, we present a Raman spectroscopy-based Process Analytical Technology (PAT) tool developed on batch and fed-batch precipitation experiments of Hepatitis B core Antigen VLPs. We conducted small-scale precipitation experiments providing a diversified data set with varying precipitation dynamics and backgrounds induced by initial dilution or spiking of clarified Escherichia coli-derived lysates. For the Raman spectroscopy data, various preprocessing operations were systematically combined allowing the identification of a preprocessing pipeline, which proved to effectively eliminate initial lysate composition variations as well as most interferences attributed to precipitates and the precipitant present in solution. The calibrated partial least squares models seamlessly predicted the precipitant concentration with R2 of 0.98 and 0.97 in batch and fed-batch experiments, respectively, and captured the observed precipitation trends with R2 of 0.74 and 0.64. Although the resolution of fine differences between experiments was limited due to the observed non-linear relationship between spectral data and the VLP concentration, this study provides a foundation for employing Raman spectroscopy as a PAT sensor for monitoring VLP precipitation processes with the potential to extend its applicability to other phase-behavior dependent processes or molecules.

show abstract

“…In recent work, using an insect-based baculovirus system, the PAT platform has been utilized to produce rabies virus-like particles. During the production process, the quantification of all stages of growth and other biochemical parameters were monitored via on-line and off-line Raman spectroscopy in association with a multivariate computational model for data analysis [ 156 ].…”

Section: Emergent Technologies In Upstream Process Developmentmentioning

confidence: 99%

Challenges and Opportunities in the Process Development of Chimeric Vaccines

Chauhan,

Khasa

2023

Vaccines

View full text Add to dashboard Cite

Vaccines are integral to human life to protect them from life-threatening diseases. However, conventional vaccines often suffer limitations like inefficiency, safety concerns, unavailability for non-culturable microbes, and genetic variability among pathogens. Chimeric vaccines combine multiple antigen-encoding genes of similar or different microbial strains to protect against hyper-evolving drug-resistant pathogens. The outbreaks of dreadful diseases have led researchers to develop economical chimeric vaccines that can cater to a large population in a shorter time. The process development begins with computationally aided omics-based approaches to design chimeric vaccines. Furthermore, developing these vaccines requires optimizing upstream and downstream processes for mass production at an industrial scale. Owing to the complex structures and complicated bioprocessing of evolving pathogens, various high-throughput process technologies have come up with added advantages. Recent advancements in high-throughput tools, process analytical technology (PAT), quality-by-design (QbD), design of experiments (DoE), modeling and simulations, single-use technology, and integrated continuous bioprocessing have made scalable production more convenient and economical. The paradigm shift to innovative strategies requires significant attention to deal with major health threats at the global scale. This review outlines the challenges and emerging avenues in the bioprocess development of chimeric vaccines.

show abstract

Biochemical monitoring throughout all stages of rabies virus-like particles production by Raman spectroscopy using global models

Cited by 11 publications

References 37 publications

Monitoring by Raman spectroscopy of rabies virus‐like particles production since the initial development stages

Monitoring by Raman spectroscopy of rabies virus‐like particles production since the initial development stages

Raman-based PAT for VLP precipitation: systematic data diversification and preprocessing pipeline identification

Challenges and Opportunities in the Process Development of Chimeric Vaccines

Contact Info

Product

Resources

About