DENV type 3 Envelope Protein [His] (DAG3060)

Recombinant DENV type 3 Envelope Protein from Insect cells [His]

Product Overview
Dengue Virus Type 3, Envelope Protein (a.a. 281-673), Recombinant. Contains Histidine tag.
Nature
Recombinant
Tag/Conjugate
His
Alternative Names
DENV; Dengue Virus Envelope Protein; DENV Envelope Protein; Dengue virus; DENV-3 E Protein; DENV-3
Procedure
5 mM EDTA
Purity
95% pure (determined by SDS-PAGE). Purified by using immobilized metal-chelate affinity chromatography.
Format
Liquid
Buffer
PBS, pH7.4
Preservative
0.1% Thimerosal, 1 μg/mL of Leupeptin, Aprotinin and Pepstatin A.
Storage
2-8°C short term, -20°C long term
Introduction
Dengue virus (DENV) is an enveloped, single-stranded, positive-sense RNA virus that includes four related but distinct serotypes (DENV1, 2, 3, and 4). It encodes three structural proteins (capsid, membrane and envelope) and seven non-structural proteins (NS1,-2a, -2b, -3, -4a, -4b and -5). The envelope (E) glycoprotein mediates virion attachment to the receptor and fusion of the virus envelope with the target cell membrane. The recombinant E protein can be used in diagnostic assays for the detection of either primary or secondary dengue infection, to overcome safety issues associated with the use of whole virus.
Keywords
DENV; Dengue Virus Envelope Protein; DENV Envelope Protein; Dengue virus; DENV-3 E Protein; DENV-3

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References


Dengue Viruses Are Enhanced by Distinct Populations of Serotype Cross-Reactive Antibodies in Human Immune Sera

PLOS PATHOGENS

Authors: de Alwis, Ruklanthi; Williams, Katherine L.; Schmid, Michael A.; Lai, Chih-Yun; Patel, Bhumi; Smith, Scott A.; Crowe, James E.; Wang, Wei-Kung; Harris, Eva; de Silva, Aravinda M.

Dengue viruses (DENV) are mosquito-borne flaviviruses of global importance. DENV exist as four serotypes, DENV1-DENV4. Following a primary infection, individuals produce DENV-specific antibodies that bind only to the serotype of infection and other antibodies that cross-react with two or more serotypes. People exposed to a secondary DENV infection with another serotype are at greater risk of developing more severe forms of dengue disease. The increased risk of severe dengue in people experiencing repeat DENV infections appear to be due, at least in part, to the ability of pre-existing serotype cross-reactive antibodies to form virus-antibody complexes that can productively infect Fc gamma receptor-bearing target cells. While the theory of antibody-dependent enhancement (ADE) is supported by several human and small animal model studies, the specific viral antigens and epitopes recognized by enhancing human antibodies after natural infections have not been fully defined. We used antibody-depletion techniques to remove DENV-specific antibody sub-populations from primary DENV-immune human sera. The effects of removing specific antibody populations on ADE were tested both in vitro using K562 cells and in vivo using the AG129 mouse model. Removal of serotype cross-reactive antibodies ablated enhancement of heterotypic virus infection in vitro and antibody-enhanced mortality in vivo. Further depletion studies using recombinant viral antigens showed that although the removal of DENV E-specific antibodies using recombinant E (rE) protein resulted in a partial reduction in DENV enhancement, there was a significant residual enhancement remaining. Competition ADE studies using prM-specific Fab fragments in human immune sera showed that both rE-specific and prM-specific antibodies in primary DENV-immune sera significantly contribute to enhancement of heterotypic DENV infection in vitro. Identification of the targets of DENV-enhancing antibodies should contribute to the development of safe, non-enhancing vaccines against dengue.

Production of tetravalent dengue virus envelope protein domain III based antigens in lettuce chloroplasts and immunologic analysis for future oral vaccine development

PLANT BIOTECHNOLOGY JOURNAL

Authors: van Eerde, Andre; Gottschamel, Johanna; Bock, Ralph; Hansen, Kristine Eraker Aasland; Munang'andu, Hetron Mweemba; Daniell, Henry; Clarke, Jihong Liu

Dengue fever is a mosquito (Aedes aegypti) -transmitted viral disease that is endemic in more than 125 countries around the world. There are four serotypes of the dengue virus (DENV 1-4) and a safe and effective dengue vaccine must provide protection against all four serotypes. To date, the first vaccine, Dengvaxia (CYD-TDV), is available after many decades' efforts, but only has moderate efficacy. More effective and affordable vaccines are hence required. Plants offer promising vaccine production platforms and food crops offer additional advantages for the production of edible human and animal vaccines, thus eliminating the need for expensive fermentation, purification, cold storage and sterile delivery. Oral vaccines can elicit humoural and cellular immunity via both the mucosal and humoral immune systems. Here, we report the production of tetravalent EDIII antigen (EDIII-1-4) in stably transformed lettuce chloroplasts. Transplastomic EDIII-1-4-expressing lettuce lines were obtained and homoplasmy was verified by Southern blot analysis. Expression of EDIII-1-4 antigens was demonstrated by immunoblotting, with the EDIII-1-4 antigen accumulating to 3.45% of the total protein content. Immunological assays in rabbits showed immunogenicity of EDIII-1-4. Our in vitro gastrointestinal digestion analysis revealed that EDIII-1-4 antigens are well protected when passing through the oral and gastric digestion phases but underwent degradation during the intestinal phase. Our results demonstrate that lettuce chloroplast engineering is a promising approach for future production of an affordable oral dengue vaccine.

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