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Optimizing the Production of gp145, an HIV-1 Envelope Glycoprotein Vaccine Candidate and Its Encapsulation in Guanosine Microparticles

We have developed a pipeline to express, purify, and characterize HIV envelope protein (Env) gp145 from Chinese hamster ovary cells, to accelerate the production of a promising vaccine candidate. First in shake flasks, then in bioreactors, we optimized the growth conditions. By adjusting the pH to 6...

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Detalles Bibliográficos
Autores principales: Akamine, Pearl, González-Feliciano, José A., Almodóvar, Ruth, Morell, Gloriner, Rivera, Javier, Capó-Vélez, Coral M., Delgado-Vélez, Manuel, Prieto-Costas, Luis, Madera, Bismark, Eichinger, Daniel, Pino, Ignacio, Rivera, José H., Ortiz-Ubarri, José, Rivera, José M., Baerga-Ortiz, Abel, Lasalde-Dominicci, José A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10221277/
https://www.ncbi.nlm.nih.gov/pubmed/37243079
http://dx.doi.org/10.3390/vaccines11050975
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author Akamine, Pearl
González-Feliciano, José A.
Almodóvar, Ruth
Morell, Gloriner
Rivera, Javier
Capó-Vélez, Coral M.
Delgado-Vélez, Manuel
Prieto-Costas, Luis
Madera, Bismark
Eichinger, Daniel
Pino, Ignacio
Rivera, José H.
Ortiz-Ubarri, José
Rivera, José M.
Baerga-Ortiz, Abel
Lasalde-Dominicci, José A.
author_facet Akamine, Pearl
González-Feliciano, José A.
Almodóvar, Ruth
Morell, Gloriner
Rivera, Javier
Capó-Vélez, Coral M.
Delgado-Vélez, Manuel
Prieto-Costas, Luis
Madera, Bismark
Eichinger, Daniel
Pino, Ignacio
Rivera, José H.
Ortiz-Ubarri, José
Rivera, José M.
Baerga-Ortiz, Abel
Lasalde-Dominicci, José A.
author_sort Akamine, Pearl
collection PubMed
description We have developed a pipeline to express, purify, and characterize HIV envelope protein (Env) gp145 from Chinese hamster ovary cells, to accelerate the production of a promising vaccine candidate. First in shake flasks, then in bioreactors, we optimized the growth conditions. By adjusting the pH to 6.8, we increased expression levels to 101 mg/L in a 50 L bioreactor, nearly twice the previously reported titer value. A battery of analytical methods was developed in accordance with current good manufacturing practices to ensure a quality biopharmaceutical. Imaged capillary isoelectric focusing verified proper glycosylation of gp145; dynamic light scattering confirmed the trimeric arrangement; and bio-layer interferometry and circular dichroism analysis demonstrated native-like properties (i.e., antibody binding and secondary structure). MALDI-TOF mass spectrometry was used as a multi-attribute platform for accurate mass determination, glycans analysis, and protein identification. Our robust analysis demonstrates that our gp145 product is very similar to a reference standard and emphasizes the importance of accurate characterization of a highly heterogeneous immunogen for the development of an effective vaccine. Finally, we present a novel guanosine microparticle with gp145 encapsulated and displayed on its surface. The unique properties of our gp145 microparticle make it amenable to use in future preclinical and clinical trials.
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spelling pubmed-102212772023-05-28 Optimizing the Production of gp145, an HIV-1 Envelope Glycoprotein Vaccine Candidate and Its Encapsulation in Guanosine Microparticles Akamine, Pearl González-Feliciano, José A. Almodóvar, Ruth Morell, Gloriner Rivera, Javier Capó-Vélez, Coral M. Delgado-Vélez, Manuel Prieto-Costas, Luis Madera, Bismark Eichinger, Daniel Pino, Ignacio Rivera, José H. Ortiz-Ubarri, José Rivera, José M. Baerga-Ortiz, Abel Lasalde-Dominicci, José A. Vaccines (Basel) Article We have developed a pipeline to express, purify, and characterize HIV envelope protein (Env) gp145 from Chinese hamster ovary cells, to accelerate the production of a promising vaccine candidate. First in shake flasks, then in bioreactors, we optimized the growth conditions. By adjusting the pH to 6.8, we increased expression levels to 101 mg/L in a 50 L bioreactor, nearly twice the previously reported titer value. A battery of analytical methods was developed in accordance with current good manufacturing practices to ensure a quality biopharmaceutical. Imaged capillary isoelectric focusing verified proper glycosylation of gp145; dynamic light scattering confirmed the trimeric arrangement; and bio-layer interferometry and circular dichroism analysis demonstrated native-like properties (i.e., antibody binding and secondary structure). MALDI-TOF mass spectrometry was used as a multi-attribute platform for accurate mass determination, glycans analysis, and protein identification. Our robust analysis demonstrates that our gp145 product is very similar to a reference standard and emphasizes the importance of accurate characterization of a highly heterogeneous immunogen for the development of an effective vaccine. Finally, we present a novel guanosine microparticle with gp145 encapsulated and displayed on its surface. The unique properties of our gp145 microparticle make it amenable to use in future preclinical and clinical trials. MDPI 2023-05-12 /pmc/articles/PMC10221277/ /pubmed/37243079 http://dx.doi.org/10.3390/vaccines11050975 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Akamine, Pearl
González-Feliciano, José A.
Almodóvar, Ruth
Morell, Gloriner
Rivera, Javier
Capó-Vélez, Coral M.
Delgado-Vélez, Manuel
Prieto-Costas, Luis
Madera, Bismark
Eichinger, Daniel
Pino, Ignacio
Rivera, José H.
Ortiz-Ubarri, José
Rivera, José M.
Baerga-Ortiz, Abel
Lasalde-Dominicci, José A.
Optimizing the Production of gp145, an HIV-1 Envelope Glycoprotein Vaccine Candidate and Its Encapsulation in Guanosine Microparticles
title Optimizing the Production of gp145, an HIV-1 Envelope Glycoprotein Vaccine Candidate and Its Encapsulation in Guanosine Microparticles
title_full Optimizing the Production of gp145, an HIV-1 Envelope Glycoprotein Vaccine Candidate and Its Encapsulation in Guanosine Microparticles
title_fullStr Optimizing the Production of gp145, an HIV-1 Envelope Glycoprotein Vaccine Candidate and Its Encapsulation in Guanosine Microparticles
title_full_unstemmed Optimizing the Production of gp145, an HIV-1 Envelope Glycoprotein Vaccine Candidate and Its Encapsulation in Guanosine Microparticles
title_short Optimizing the Production of gp145, an HIV-1 Envelope Glycoprotein Vaccine Candidate and Its Encapsulation in Guanosine Microparticles
title_sort optimizing the production of gp145, an hiv-1 envelope glycoprotein vaccine candidate and its encapsulation in guanosine microparticles
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10221277/
https://www.ncbi.nlm.nih.gov/pubmed/37243079
http://dx.doi.org/10.3390/vaccines11050975
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