A recombinant gp145 Env glycoprotein from HIV-1 expressed in two different cell lines: Effects on glycosylation and antigenicity.

Autor: González-Feliciano JA; Molecular Sciences Research Center Inc., University of Puerto Rico, San Juan, Puerto Rico., Akamine P; Molecular Sciences Research Center Inc., University of Puerto Rico, San Juan, Puerto Rico., Capó-Vélez CM; Molecular Sciences Research Center Inc., University of Puerto Rico, San Juan, Puerto Rico., Delgado-Vélez M; Molecular Sciences Research Center Inc., University of Puerto Rico, San Juan, Puerto Rico., Dussupt V; U.S. Military HIV Research Program, Walter Reed Army Institute of Research, Silver Spring, Maryland, United States of America.; Henry M. Jackson Foundation for the Advancement of Military Medicine, Bethesda, Maryland, United States of America., Krebs SJ; U.S. Military HIV Research Program, Walter Reed Army Institute of Research, Silver Spring, Maryland, United States of America.; Henry M. Jackson Foundation for the Advancement of Military Medicine, Bethesda, Maryland, United States of America., Wojna V; Division of Neurology, Internal Medicine Department and NeuroHIV Research Program, University of Puerto Rico, Medical Sciences Campus, San Juan, Puerto Rico., Polonis VR; U.S. Military HIV Research Program, Walter Reed Army Institute of Research, Silver Spring, Maryland, United States of America., Baerga-Ortiz A; Molecular Sciences Research Center Inc., University of Puerto Rico, San Juan, Puerto Rico.; Department of Biochemistry, University of Puerto Rico, Medical Sciences Campus, San Juan, Puerto Rico., Lasalde-Dominicci JA; Molecular Sciences Research Center Inc., University of Puerto Rico, San Juan, Puerto Rico.; Department of Biology, University of Puerto Rico, Rio Piedras Campus, San Juan, Puerto Rico.
Jazyk: angličtina
Zdroj: PloS one [PLoS One] 2020 Jun 19; Vol. 15 (6), pp. e0231679. Date of Electronic Publication: 2020 Jun 19 (Print Publication: 2020).
DOI: 10.1371/journal.pone.0231679
Abstrakt: The envelope glycoprotein (Env) of the human immunodeficiency virus (HIV), has been the primary target for the development of a protective vaccine against infection. The extensive N-linked glycosylation on Env is an important consideration as it may affect efficacy, stability, and expression yields. The expression host has been shown to influence the extent and type of glycosylation that decorates the protein target. Here, we report the glycosylation profile of the candidate subtype C immunogen CO6980v0c22 gp145, which is currently in Phase I clinical trials, produced in two different host cells: CHO-K1 and Expi293F. The amino acid sequence for both glycoproteins was confirmed to be identical by peptide mass fingerprinting. However, the isoelectric point of the proteins differed; 4.5-5.5 and 6.0-7.0 for gp145 produced in CHO-K1 and Expi293F, respectively. These differences in pI were eliminated by enzymatic treatment with sialidase, indicating a large difference in the incorporation of sialic acid between hosts. This dramatic difference in the number of sialylated glycans between hosts was confirmed by analysis of PNGase F-released glycans using MALDI-ToF MS. These differences in glycosylation, however, did not greatly translate into differences in antibody recognition. Biosensor assays showed that gp145 produced in CHO-K1 had similar affinity toward the broadly neutralizing antibodies, 2G12 and PG16, as the gp145 produced in Expi293F. Additionally, both immunogens showed the same reactivity against plasma of HIV-infected patients. Taken together, these results support the notion that there are sizeable differences in the glycosylation of Env depending on the expression host. How these differences translate to vaccine efficacy remains unknown.
Competing Interests: The authors have declared that no competing interests exist.
Databáze: MEDLINE
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