Development of an enzyme-linked immunospot assay for determination of rotavirus infectivity

Li, Tingdong; Lin, Haijun; Yu, Linqi; Xue, Miaoge; Ge, Shengxiang; Zhao, Qinjian; Zhang, Jun; Xia, Ningshao

doi:10.1016/j.jviromet.2014.08.012

Cited by 16 publications

(13 citation statements)

References 42 publications

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“…Kafi et al 2008 ; Virel et al 2010 ; Zhong et al 2011 ; Ahammad et al 2011 ). In recent years however, a considerable number of studies also dealt with the detection of other molecules, such as glucose (Alonso-Lomillo et al 2005 ), ethanol (Azevedo et al 2005 ), DNA and RNA (Fan et al 2013 ; Tran et al 2014 ; Saikrishnan et al 2014 ), l -phenylalanine (Kubota et al 2013 ), citrinin (Zachetti et al 2013 ), pyrogallol and hydroquinone (Raghu et al 2013 ), phenols (Kafi and Chen 2009 ; Liu et al 2011 ), the milk allergen β-lactoglobulin (Ruiz-Valdepeñas Montiel et al 2015 ), rotavirus titers (Li et al 2014 ), and tumor markers (Chen et al 2009 ; Kim et al 2014b ; Patris et al 2014 ) via H 2 O 2 . Notably, studies on HRP applications of medical relevance are currently particularly prevalent.…”

Section: Impact Of Recombinant Technology On Traditional and Future Hmentioning

confidence: 99%

An updated view on horseradish peroxidases: recombinant production and biotechnological applications

Krainer

Glieder

2015

Appl Microbiol Biotechnol

191

133

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Horseradish peroxidase has been the subject of scientific research for centuries. It has been used exhaustively as reporter enzyme in diagnostics and histochemistry and still plays a major role in these applications. Numerous studies have been conducted on the role of horseradish peroxidase in the plant and its catalytic mechanism. However, little progress has been made in its recombinant production. Until now, commercial preparations of horseradish peroxidase are still isolated from plant roots. These preparations are commonly mixtures of various isoenzymes of which only a small fraction has been described so far. The composition of isoenzymes in these mixed isolates is subjected to uncontrollable environmental conditions. Nowadays, horseradish peroxidase regains interest due to its broad applicability in the fields of medicine, life sciences, and biotechnology in cancer therapy, biosensor systems, bioremediation, and biocatalysis. These medically and commercially relevant applications, the recent discovery of new natural isoenzymes with different biochemical properties, as well as the challenges in recombinant production render this enzyme particularly interesting for future biotechnological solutions. Therefore, we reviewed previous studies as well as current developments with biotechnological emphasis on new applications and the major remaining biotechnological challenge—the efficient recombinant production of horseradish peroxidase enzymes.

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Section: Impact Of Recombinant Technology On Traditional and Future Hmentioning

confidence: 99%

An updated view on horseradish peroxidases: recombinant production and biotechnological applications

Krainer

Glieder

2015

Appl Microbiol Biotechnol

191

133

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“…46 The infectious titer of rotavirus was determined by an enzyme-linked immune-spot (Elispot) assay. 47 When used as antigen for immunization, the virus was inactivated by 1% formalin treatment at 37 C for 72 h, and then the excess formalin was neutralized with 0.05% sodium bisulfite for 30 min followed by dialysis against PBS; the inactivation was verified by a TCID 50 assay to demonstrate the lack of a cytopathic effect.…”

Section: Virus Stockmentioning

confidence: 99%

“…The neutralizing activities of the sera from mice immunized with recombinant proteins were determined by Elispot assay as previously described. 47 Briefly, 100 ml aliquots of 2-fold serially diluted serum were mixed with equal volumes of trypsinactivated rotavirus LLR; after neutralization, 100 ml of each serum-rotavirus mixture was added to MA104 cells in 96-well microplates. After 14 h of incubation, reductions in rotavirus infectivity were measured by Elispot.…”

Section: Micro-neutralization Assaymentioning

confidence: 99%

Immunogenicity and protective efficacy of rotavirus VP8*fused to cholera toxin B subunit in a mouse model

Xue

Jia

et al. 2016

Human Vaccines & Immunotherapeutics

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In attempts to develop recombinant subunit vaccines against rotavirus disease, it was previously shown that the N-terminal truncated VP8* protein, VP8-1 (aa26-231), is a good vaccine candidate when used for immunization in combination with Freund's adjuvant. However, this protein stimulated only weak immune response when aluminum hydroxide was used as an adjuvant. In this study, the nontoxic B subunit of cholera toxin (CTB) was employed as intra-molecular adjuvant to improve the immunogenicity of VP8-1. Both, the N-terminal and C-terminal fusion proteins, were purified to homogeneity, at which stage they formed pentamers, and showed significantly higher immunogenicity and protective efficacy than a VP8-1/aluminum hydroxide mixture in a mouse model. Compared to VP8-1-CTB, CTB-VP8-1 showed higher binding activity to both, GM1 and the conformation sensitive neutralizing monoclonal antibodies specific to VP8. More importantly, CTB-VP8-1 elicited higher titers of neutralizing antibodies and conferred higher protective efficacy than VP8-1-CTB. Therefore, the protein CTB-VP8-1, with enhanced immunogenicity and immunoprotectivity, could be considered as a viable candidate for further development of an alternative, replication-incompetent, parenterally administered vaccine against rotavirus disease.

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“…was cultured in MA-104 cells (ATCC ® CRL2378.1) (ATCC, Manassas, VA) as described previously [35]. The infectious titer of rotavirus was determined by an enzyme-linked immune-spot (Elispot) assay [36].…”

Section: Viruses and Cell Culturementioning

confidence: 99%

“…The neutralizing antibody titer of each serum sample was determined by an Elispot-based micro-neutralization assay [36]. Briefly, an aliquot of 500 l of 2-fold serially diluted serum samples were mixed with an equal volume of trypsin-activated rotavirus LLR and were incubated at 37 • C for 1 h, then 200 l of the mixture was added to a MA-104 monolayer (two replicates) and was incubated for 14 h at 37 • C before detection.…”

Section: Micro-neutralization Assaymentioning

confidence: 99%

Characterization and protective efficacy in an animal model of a novel truncated rotavirus VP8 subunit parenteral vaccine candidate

Xue

Che

et al. 2015

Vaccine

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Development of an enzyme-linked immunospot assay for determination of rotavirus infectivity

Cited by 16 publications

References 42 publications

An updated view on horseradish peroxidases: recombinant production and biotechnological applications

An updated view on horseradish peroxidases: recombinant production and biotechnological applications

Immunogenicity and protective efficacy of rotavirus VP8*fused to cholera toxin B subunit in a mouse model

Characterization and protective efficacy in an animal model of a novel truncated rotavirus VP8 subunit parenteral vaccine candidate

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