Recombinant dengue virus-like particles from Pichia pastoris: efficient production and immunological properties

Abstract

The envelope glycoprotein (E) of flavivirus is the major structural protein on the surface of the mature virions. The complexes of premembrane (prM) and E play important roles in virus assembly and fusion modulation and in potential immunity-inducing vaccines. In the present study, the cDNA encoding prM and E proteins of dengue virus type 2 (DENV-2) was subcloned into the pGAPZalphaA vector and further integrated into the genome of Pichia pastoris under the control of the glyceraldehyde-3-phosphate dehydrogenase (GAP) constitutive promoter. The high-level constitutive expression of recombinant E antigen was achieved in P. pastoris. Both the cell lysate and the culture supernatant, examined by electron microscopy, were found to contain DENV-2 virus-like particles (VLPs) with diameters of about 30 nm. After immunization of BALB/c mice, the VLPs exhibited similar efficacies as inactivated virus in terms of antibody induction and neutralization titer. These results suggest that recombinant DENV VLPs can be efficiently produced in the GAP promoter-based P. pastoris expression system. This system may be useful for the development of effective and economic dengue subunit vaccine.

Retracted: Immunological properties reveal the monovalent and bivalent recombinant dengue virus‐like particles as candidate vaccine for dengue

“…P. pastoris yeast cells characterized by tolerance, were found to the most economic expression systems for vaccine development (21) and GAP promoter was successfully used in expressing large amount of HBsAg in yeast cells (22). The GAP promoter-based P. pastoris expression system with the signal peptide of prM has been determined to be safe and stable for constitutive and effective production of DENV-2 VLPs (23). In contrast, we applied α-factor secretion signal in the GAP promoter-P. pastoris expression system and obtained high-level production of DENV-1 VLPs and DENV-2 VLPs.…”

An Alphavirus Vector-Based Tetravalent Dengue Vaccine Induces a Rapid and Protective Immune Response in Macaques That Differs Qualitatively from Immunity Induced by Live Virus Infection

“…In this study, we compared the antibody responses induced by infectious DENV virions, recombinant DENV subviral particles (prME), and soluble E dimers (E85). Coexpression of prM and E of dengue virus and other flaviviruses using heterologous expression systems results in the assembly of recombinant subviral particles (74)(75)(76)(77)(78)(79). These subviral particles are smaller than dengue virus virions, have an icosahedral symmetry of T ϭ 1 instead of T ϭ 3, and may display E proteins with different organizations.…”

Naturally mutated envelope protein domain I of Chinese B dengue virus attenuated human dendritic cell maturation

“…The envelope protein is the viral structural glycoprotein that resides on the surface of mature virions. It plays important roles in virus assembly, cell attachment and membrane fusion and could potentially be used to generate immunity-inducing vaccines [24]. The E protein folds into three distinct structural or functional domains.…”