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

Autor: Akamine P; Clinical Bioreagent Center, Molecular Sciences Research Center, University of Puerto Rico, San Juan 00926, Puerto Rico., González-Feliciano JA; Clinical Bioreagent Center, Molecular Sciences Research Center, University of Puerto Rico, San Juan 00926, Puerto Rico., Almodóvar R; CDI Laboratories, Mayagüez 00680, Puerto Rico., Morell G; CDI Laboratories, Mayagüez 00680, Puerto Rico., Rivera J; CDI Laboratories, Mayagüez 00680, Puerto Rico., Capó-Vélez CM; Clinical Bioreagent Center, Molecular Sciences Research Center, University of Puerto Rico, San Juan 00926, Puerto Rico., Delgado-Vélez M; Clinical Bioreagent Center, Molecular Sciences Research Center, University of Puerto Rico, San Juan 00926, Puerto Rico.; Department of Biology, Río Piedras Campus, University of Puerto Rico, San Juan 00931, Puerto Rico., Prieto-Costas L; Department of Chemistry, Río Piedras Campus, University of Puerto Rico, San Juan 00925, Puerto Rico., Madera B; Clinical Bioreagent Center, Molecular Sciences Research Center, University of Puerto Rico, San Juan 00926, Puerto Rico.; Department of Biology, Río Piedras Campus, University of Puerto Rico, San Juan 00931, Puerto Rico., Eichinger D; CDI Laboratories, Mayagüez 00680, Puerto Rico., Pino I; CDI Laboratories, Mayagüez 00680, Puerto Rico., Rivera JH; CDI Laboratories, Mayagüez 00680, Puerto Rico., Ortiz-Ubarri J; Department of Computer Sciences, Río Piedras Campus, University of Puerto Rico, San Juan 00925, Puerto Rico., Rivera JM; Department of Chemistry, Río Piedras Campus, University of Puerto Rico, San Juan 00925, Puerto Rico., Baerga-Ortiz A; Clinical Bioreagent Center, Molecular Sciences Research Center, University of Puerto Rico, San Juan 00926, Puerto Rico.; Department of Biochemistry, Medical Sciences Campus, University of Puerto Rico, San Juan 00936, Puerto Rico., Lasalde-Dominicci JA; Clinical Bioreagent Center, Molecular Sciences Research Center, University of Puerto Rico, San Juan 00926, Puerto Rico.; Department of Biology, Río Piedras Campus, University of Puerto Rico, San Juan 00931, Puerto Rico.; Department of Chemistry, Río Piedras Campus, University of Puerto Rico, San Juan 00925, Puerto Rico.; Institute of Neurobiology, Medical Sciences Campus, University of Puerto Rico, San Juan 00901, Puerto Rico.
Jazyk: angličtina
Zdroj: Vaccines [Vaccines (Basel)] 2023 May 12; Vol. 11 (5). Date of Electronic Publication: 2023 May 12.
DOI: 10.3390/vaccines11050975
Abstrakt: 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.
Databáze: MEDLINE
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