Production of recombinant NS1 protein and its possible use in encephalitic flavivirus differential diagnosis

Lorch, Matías Sebastián; Collado, María Soledad; Argüelles, Marcelo Horacio; Rota, Rosana P.; Spinsanti, Lorena; Lozano, Mario Enrique; Goñi, Sandra Elizabeth

doi:10.1016/j.pep.2018.08.008

Cited by 15 publications

(5 citation statements)

References 34 publications

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“…Different strategies for the production and purification of flavivirus rNS1 proteins and their use in the development of NS1-based ELISA assays have been reported, based on rNS1 proteins produced in systems that are prone to problems of protein stability and lack the proper folding and post-translational modifications [35][36][37]. In the present study, we report a strategy for the production and purification of rNS1 proteins of TBEV, WNV, ZIKV and all DENV serotypes from the supernatant of transiently transfected HEK293T cells.…”

Section: Discussionmentioning

confidence: 99%

Comparative specificity and sensitivity of NS1-based serological assays for the detection of flavivirus immune response

et al. 2020

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Flaviviruses are relevant animal and human pathogens of increasing importance worldwide. The similarities of the initial clinical symptoms and the serological cross-reactivity of viral structural antigens make a laboratory diagnosis of flavivirus infection problematic. The main aim of the present study was the comparative specificity and sensitivity analysis of the nonstructural protein NS1 as an antigen to detect flavivirus antibodies in sera from exposed individuals. A strategy for the purification of native recombinant non-structural protein 1 of representative flaviviruses including tick-borne encephalitis, West Nile, Zika and dengue virus was developed. The immunological properties of the purified antigens were analyzed using sera of immunized mice and of infected individuals in comparison with standard commercial assays. Recombinant NS1 protein was confirmed as a valuable option for the detection of flavivirus antibodies with reduced cross-reactivity and high sensitivity offering additional advantages for the detection of vaccine breakthrough cases.

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Section: Discussionmentioning

confidence: 99%

Comparative specificity and sensitivity of NS1-based serological assays for the detection of flavivirus immune response

et al. 2020

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“…However, several other factors may contribute to the inclusion bodies formation [63] and it is very common during overexpression of heterologous genes in E. coli , particularly from animal origin. Although the biological activity of the protein in this state is impaired, some studies show that insoluble proteins can successfully be used to produce polyclonal antibodies [64,65,66]. In addition, the inclusion bodies may represent some advantages since they are less vulnerable to degradation and may remain longer in tissues, avoiding their fast clearance, which could, in theory, require fewer boosters or even the necessity of using adjuvants.…”

Section: Discussionmentioning

confidence: 99%

Design and Production of a Recombinant Hybrid Toxin to Raise Protective Antibodies against Loxosceles Spider Venom

Pal

Lhal

Colombini

et al. 2019

Toxins

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Human accidents with spiders of the genus Loxosceles are an important health problem affecting thousands of people worldwide. Patients evolve to severe local injuries and, in many cases, to systemic disturbances as acute renal failure, in which cases antivenoms are considered to be the most effective treatment. However, for antivenom production, the extraction of the venom used in the immunization process is laborious and the yield is very low. Thus, many groups have been exploring the use of recombinant Loxosceles toxins, particularly phospholipases D (PLDs), to produce the antivenom. Nonetheless, some important venom activities are not neutralized by anti-PLD antibodies. Astacin-like metalloproteases (ALMPs) are the second most expressed toxin acting on the extracellular matrix, indicating the importance of its inclusion in the antigen’s formulation to provide a better antivenom. Here we show the construction of a hybrid recombinant immunogen, called LgRec1ALP1, composed of hydrophilic regions of the PLD and the ALMP toxins from Loxosceles gaucho. Although the LgRec1ALP1 was expressed as inclusion bodies, it resulted in good yields and it was effective to produce neutralizing antibodies in mice. The antiserum neutralized fibrinogenolytic, platelet aggregation and dermonecrotic activities elicited by L. gaucho, L. laeta, and L. intermedia venoms, indicating that the hybrid recombinant antigen may be a valuable source for the production of protective antibodies against Loxosceles ssp. venoms. In addition, the hybrid recombinant toxin approach may enrich and expand the alternative antigens for antisera production for other venoms.

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“…Both antibodies to this protein and the antigen itself in the serum of an infected person may appear earlier than the immune response to protein E [14]. In addition, test systems based on the NS1 antigen are currently used for epidemiologic diagnosis, including differential diagnosis of the Saint Louis encephalitis and West Nile viruses [15].…”

Section: Introductionmentioning

confidence: 99%

Cross-Reactive Antibodies to the NS1 Protein of Omsk Hemorrhagic Fever Virus Are Absent in the Sera of Patients with Tick-Borne Encephalitis

Kravchuk,

Khlusevich,

Chicherina

et al. 2024

Preprint

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Omsk hemorrhagic fever virus (OHFV) is a member of the tick-borne encephalitis virus (TBEV) complex of the Flaviviridae family. Currently, there is no data on cross-reactivity of antibodies to the NS1 proteins of OHFV and TBEV. Such data are of major interest for monitoring viral encephalitis of unknown etiology due to the increasing geographical distribution of OHFV. In this study, recombinant OHFV NS1 protein was produced using Escherichia сoli expression system and purified. The recombinant OHFV NS1 protein was recognized by specific mice immune ascetic fluids to the native OHFV NS1 protein. Western blot analysis and ELISA of recombinant NS1 proteins of OHFV and TBEV were used to study the cross-reactivity of antibodies from immune ascites fluid obtained from OHFV-infected mice and mAbs against TBEV NS1. Anti-TBEV NS1 mouse monoclonal antibodies (mAbs) have been shown not to be cross-reactive to OHFV NS1 protein. Sera from patients with confirmed tick-borne encephalitis (TBE) were examined by ELISA using recombinant OHFV NS1 and TBEV NS1 proteins as antigens. It was shown for the first time that cross-reactive antibodies to OHFV NS1 protein were not detected in the sera of TBE patients whereas the sera contained antibodies to TBEV NS1 protein.

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Production of recombinant NS1 protein and its possible use in encephalitic flavivirus differential diagnosis

Cited by 15 publications

References 34 publications

Comparative specificity and sensitivity of NS1-based serological assays for the detection of flavivirus immune response

Comparative specificity and sensitivity of NS1-based serological assays for the detection of flavivirus immune response

Design and Production of a Recombinant Hybrid Toxin to Raise Protective Antibodies against Loxosceles Spider Venom

Cross-Reactive Antibodies to the NS1 Protein of Omsk Hemorrhagic Fever Virus Are Absent in the Sera of Patients with Tick-Borne Encephalitis

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