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

Gonzalez-Feliciano, J. A.; Akamine, P. Y.; Capo-Velez, C. M.; Delgado-Velez, M.; Dussupt, V.; Krebs, S. J.; Wojna, V.; Polonis, V. R.; Baerga-Ortiz, A.; Lasalde-Dominicci, J. A.

doi:10.1101/2020.03.31.018408

bioRxiv · 10.1101/2020.03.31.018408

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

Gonzalez-Feliciano, J. A.·Akamine, P. Y.·Capo-Velez, C. M.·Delgado-Velez, M.·Dussupt, V.·Krebs, S. J.·Wojna, V.·Polonis, V. R.·Baerga-Ortiz, A.·Lasalde-Dominicci, J. A.

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Abstract

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 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.

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Gonzalez-Feliciano, J. A., Akamine, P. Y., Capo-Velez, C. M., Delgado-Velez, M., Dussupt, V., Krebs, S. J., Wojna, V., Polonis, V. R., Baerga-Ortiz, A., Lasalde-Dominicci, J. A.. 2020-03-31. A recombinant gp145 Env glycoprotein from HIV-1 expressed in two different cell lines: effects on glycosylation and antigenicity. https://doi.org/10.1101/2020.03.31.018408

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