DENV type 2 Envelope protein [His] (DAG453)

DENV type 2 Envelope protein (aa 300 - 400) [His], recombinant protein from E. coli

Product Overview
Recombinant Dengue Type-2 Envelope Domain III (amino acids 300-400) immunodominant regions. It was expressed in E.coli. This region contains neutralizing epitopes, and receptor binding domain. Reacts with Dengue Type 2 IgG. Contains a 6-His fusion partner
Nature
Recombinant
Tag/Conjugate
His
Molecular Weight
12 kDa
Procedure
None
Purity
> 95% pure (12% PAGE (Coomassie)). Proprietary chromatographic technique.
Format
Purified, Liquid
Concentration
1.92 mg/ml
Buffer
PBS, pH 7.4
Preservative
None
Storage
2-8°C short term, -20°C long term
Introduction
Dengue virus (DENV) in one of four serotypes is the cause of dengue fever. It is a mosquito-borne single positive-stranded RNA virus of the family Flaviviridae; genus Flavivirus. All four serotypes can cause the full spectrum of disease. Its genome is about 11000 bases that codes for three structural proteins, capsid protein C, membrane protein M, envelope protein E; seven nonstructural proteins, NS1, NS2a, NS2b, NS3, NS4a, NS4b, NS5; and short non-coding regions on both the 5' and 3' ends. Further classification of each serotype into genotypes often relates to the region where particular strains are commonly found or were first found.
Antigen Description
Envelope protein E binds cell surface receptor and is involved in membrane fusion between virion and target cell. Synthesized as a homodimer with prM which acts as a chaperone for envelope protein E. After cleavage of prM, envelope protein E dissociates from small envelope protein M and homodimerizes.
Keywords
DENV; Dengue Virus Envelope Protein; DENV Envelope Protein; Dengue virus

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References


Pichia pastoris-expressed Zika virus envelope domain III on a virus-like particle platform: design, production and immunological evaluation

PATHOGENS AND DISEASE

Authors: Shanmugam, Rajgokul K.; Ramasamy, Viswanathan; Shukla, Rahul; Arora, Upasana; Swaminathan, Sathyamangalam; Khanna, Navin

Zika virus (ZIKV) is an arbovirus which shares antigenic similarity and the mosquito vector with dengue viruses (DENVs). ZIKV is a neurotropic virus capable of causing congenital neurodevelopmental birth defects. As ZIKV antibodies (Abs) can potentially enhance infection by DENVs, a preventive ZIKV vaccine must be designed to eliminate antibody dependent enhancement of infection. We developed a Zika Subunit Vaccine (ZSV) consisting of two proteins, ZS and S, in a genetically pre-determined ratio of 1:4, using the methylotrophic yeast Pichia pastoris. ZS is an in-frame fusion of ZIKV envelope domain III with the Hepatitis B virus (HBV) surface antigen, and S is the un-fused HBV surface antigen. Using specific monoclonal Abs we showed the presence of ZS and S in the co-purified material which were found to co-assemble into virus-like particles (VLPs), based on dynamic light scattering and electron microscopic analyses. These VLPs were immunogenic in BALB/c mice, eliciting Abs capable of neutralizing ZIKV reporter virus particles. Further, the VLP-induced Abs did not enhance a sub-lethal DENV-2 challenge in AG129 mice. This important safety feature, coupled to the well-documented advantage of P. pastoris expression system, warrants further exploration of ZSV VLP as a possible vaccine candidate.

Preclinical development of a dengue tetravalent recombinant subunit vaccine: Immunogenicity and protective efficacy in nonhuman primates

VACCINE

Authors: Govindarajan, Dhanasekaran; Meschino, Steven; Guan, Liming; Clements, David E.; ter Meulenc, Jan H.; Casimiro, Danilo R.; Collera, Beth-Ann G.; Bett, Andrew J.

We describe here the preclinical development of a dengue vaccine composed of recombinant subunit carboxy-truncated envelope (E) proteins (DEN-80E) for each of the four dengue serotypes. Immunogenicity and protective efficacy studies in Rhesus monkeys were conducted to evaluate monovalent and tetravalent DEN-80E vaccines formulated with ISCOMATRIX (TM) adjuvant. Three different doses and two dosing regimens (0, 1, 2 months and 0, 1, 2, and 6 months) were evaluated in these studies. We first evaluated monomeric (DEN4-80E) and dimeric (DEN4-80EZip) versions of DEN4-80E, the latter generated in an attempt to improve immunogenicity. The two antigens, evaluated at 6,20 and 100 mu g/dose formulated with ISCOMATRIX (TM) adjuvant, were equally immunogenic. A group immunized with 20 mu g DEN4-80E and Alhydrogel (TM) induced much weaker responses. When challenged with wild-type dengue type 4 virus, all animals in the 6 and 20 mu g groups and all but one in the DEN4-80EZip 100 mu g group were protected from viremia. Two out of three monkeys in the Alhydrogel (TM) group had breakthrough viremia. A similar study was conducted to evaluate tetravalent formulations at low (3, 3, 3, 6 mu g of DEN1-80E, DEN2-80E, DEN3-80E and DEN4-80E respectively), medium (10, 10, 10, 20 mu g) and high (50, 50, 50, 100 mu g) doses. All doses were comparably immunogenic and induced high titer, balanced neutralizing antibodies against all four DENV. Upon challenge with the four wild-type DENV, all animals in the low and medium dose groups were protected against viremia while two animals in the high-dose group exhibited breakthrough viremia. Our studies also indicated that a 0, 1, 2 and 6 month vaccination schedule is superior to the 0, 1, and 2 month schedule in terms of durability. Overall, the subunit vaccine was demonstrated to induce strong neutralization titers resulting in protection against viremia following challenge even 8-12 months after the last vaccine dose. (C) 2015 Elsevier Ltd. All rights reserved.

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