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Volume 14
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10.3390/v14112539
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Article

Assessing Multi-Attribute Characterization of Enveloped and Non-Enveloped Viral Particles by Capillary Electrophoresis

by
Rita P. Fernandes
1,2,†,
José M. Escandell
1,2,†,
Ana C. L. Guerreiro
1,2,
Filipa Moura
1,2,
Tiago Q. Faria
1,2,
Sofia B. Carvalho
1,2,
Ricardo J. S. Silva
1,2,
Patrícia Gomes-Alves
1,2,* and
Cristina Peixoto
1,2,*
1
iBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2780-901 Oeiras, Portugal
2
Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Avenida da República, 2780-157 Oeiras, Portugal
*
Authors to whom correspondence should be addressed.
†
These authors contributed equally to this work.
Viruses 2022, 14(11), 2539; https://doi.org/10.3390/v14112539
Submission received: 19 October 2022 / Revised: 13 November 2022 / Accepted: 15 November 2022 / Published: 17 November 2022
(This article belongs to the Section General Virology)
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Abstract

Virus-based biopharmaceutical products are used in clinical applications such as vaccines, gene therapy, and immunotherapy. However, their manufacturing remains a challenge, hampered by the lack of appropriate analytical tools for purification monitoring or characterization of the final product. This paper describes the implementation of a highly sensitive method, capillary electrophoresis (CE)-sodium dodecyl sulfate (SDS) combined with a laser-induced fluorescence (LIF) detector to monitor the impact of various bioprocess steps on the quality of different viral vectors. The fluorescence labelling procedure uses the (3-(2-furoyl) quinoline-2-carboxaldehyde dye, and the CE-SDS LIF method enables the evaluation of in-process besides final product samples. This method outperforms other analytical methods, such as SDS–polyacrylamide gel electrophoresis with Sypro Ruby staining, in terms of sensitivity, resolution, and high-throughput capability. Notably, this CE-SDS LIF method was also successfully implemented to characterize enveloped viruses such as Maraba virus and lentivirus, whose development as biopharmaceuticals is now restricted by the lack of suitable analytical tools. This method was also qualified for quantification of rAAV2 according to the International Council for Harmonisation guidelines. Overall, our work shows that CE-SDS LIF is a precise and sensitive analytical platform for in-process sample analysis and quantification of different virus-based targets, with a great potential for application in biomanufacturing.
Keywords: bioanalytical tools; capillary electrophoresis; quality attributes; virus-based therapeutics bioanalytical tools; capillary electrophoresis; quality attributes; virus-based therapeutics

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MDPI and ACS Style

Fernandes, R.P.; Escandell, J.M.; Guerreiro, A.C.L.; Moura, F.; Faria, T.Q.; Carvalho, S.B.; Silva, R.J.S.; Gomes-Alves, P.; Peixoto, C. Assessing Multi-Attribute Characterization of Enveloped and Non-Enveloped Viral Particles by Capillary Electrophoresis. Viruses 2022, 14, 2539. https://doi.org/10.3390/v14112539

AMA Style

Fernandes RP, Escandell JM, Guerreiro ACL, Moura F, Faria TQ, Carvalho SB, Silva RJS, Gomes-Alves P, Peixoto C. Assessing Multi-Attribute Characterization of Enveloped and Non-Enveloped Viral Particles by Capillary Electrophoresis. Viruses. 2022; 14(11):2539. https://doi.org/10.3390/v14112539

Chicago/Turabian Style

Fernandes, Rita P., José M. Escandell, Ana C. L. Guerreiro, Filipa Moura, Tiago Q. Faria, Sofia B. Carvalho, Ricardo J. S. Silva, Patrícia Gomes-Alves, and Cristina Peixoto. 2022. "Assessing Multi-Attribute Characterization of Enveloped and Non-Enveloped Viral Particles by Capillary Electrophoresis" Viruses 14, no. 11: 2539. https://doi.org/10.3390/v14112539

APA Style

Fernandes, R. P., Escandell, J. M., Guerreiro, A. C. L., Moura, F., Faria, T. Q., Carvalho, S. B., Silva, R. J. S., Gomes-Alves, P., & Peixoto, C. (2022). Assessing Multi-Attribute Characterization of Enveloped and Non-Enveloped Viral Particles by Capillary Electrophoresis. Viruses, 14(11), 2539. https://doi.org/10.3390/v14112539

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