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Quantitative and sensitive detection of the SARS-CoV spike protein using bispecific monoclonal antibody-based enzyme-linked immunoassay.

The severe acute respiratory syndrome coronavirus (SARS-CoV) spike protein is known to mediate receptor interaction and immune recognition and thus it is considered as a major target for vaccine design. The spike protein plays an important role in virus entry, virus receptor interactions, and virus tropism. Sensitive diagnosis of SARS is essential for the control of the disease in humans. Recombinant SARS-CoV S1 antigen was produced and purified for the development of monoclonal and bi-specific monoclonal antibodies. The hybridomas secreting anti-S1 antibodies, F26G18 and P136.8D12, were fused respectively with the YP4 hybridoma to generate quadromas. The sandwich ELISA was formed by using F26G18 as a coating antibody and biotinylated F26G18 as a detection antibody with a detection limit of 0.037μg/ml (p<0.02). The same detection limit was found with P136.8D12 as a coating antibody and biotinylated F26G18 as a detection antibody. The sensitivity was improved (detection limit of 0.019μg/ml), however, when using bi-specific monoclonal antibody (F157) as the detection antibody. In conclusion, the method described in this study allows sensitive detection of a recombinant SARS spike protein by sandwich ELISA with bi-specific monoclonal antibody and could be used for the diagnosis of patients suspected with SARS.

2774 related Products with: Quantitative and sensitive detection of the SARS-CoV spike protein using bispecific monoclonal antibody-based enzyme-linked immunoassay.

Leptin ELISA Kit, Human A Anti C Reactive Protein A Rabbit Anti-SARS-Associat Rabbit Anti-SARS-Associat Rabbit Anti-SARS-Associat Anti CEL Monoclonal Antib Anti AGE 3 Monoclonal Ant anti GFP antibody, rat mo Anti-SARS Spike Protein I Proteinase 3 Antibody Mou Rabbit Anti-F1 protein (Y Rabbit Anti-F1 protein (Y

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Dendritic cell targeted chitosan nanoparticles for nasal DNA immunization against SARS CoV nucleocapsid protein.

This work investigates the formulation and in vivo efficacy of dendritic cell (DC) targeted plasmid DNA loaded biotinylated chitosan nanoparticles for nasal immunization against nucleocapsid (N) protein of severe acute respiratory syndrome coronavirus (SARS-CoV) as antigen. The induction of antigen-specific mucosal and systemic immune response at the site of virus entry is a major challenge for vaccine design. Here, we designed a strategy for noninvasive receptor mediated gene delivery to nasal resident DCs. The pDNA loaded biotinylated chitosan nanoparticles were prepared using a complex coacervation process and characterized for size, shape, surface charge, plasmid DNA loading and protection against nuclease digestion. The pDNA loaded biotinylated chitosan nanoparticles were targeted with bifunctional fusion protein (bfFp) vector for achieving DC selective targeting. The bfFp is a recombinant fusion protein consisting of truncated core-streptavidin fused with anti-DEC-205 single chain antibody (scFv). The core-streptavidin arm of fusion protein binds with biotinylated nanoparticles, while anti-DEC-205 scFv imparts targeting specificity to DC DEC-205 receptor. We demonstrate that intranasal administration of bfFp targeted formulations along with anti-CD40 DC maturation stimuli enhanced magnitude of mucosal IgA as well as systemic IgG against N protein. The strategy led to the detection of augmented levels of N protein specific systemic IgG and nasal IgA antibodies. However, following intranasal delivery of naked pDNA no mucosal and systemic immune responses were detected. A parallel comparison of targeted formulations using intramuscular and intranasal routes showed that the intramuscular route is superior for induction of systemic IgG responses compared with the intranasal route. Our results suggest that targeted pDNA delivery through a noninvasive intranasal route can be a strategy for designing low-dose vaccines.

2411 related Products with: Dendritic cell targeted chitosan nanoparticles for nasal DNA immunization against SARS CoV nucleocapsid protein.

Rabbit Anti-SARS Virus Nu Rabbit Anti-SARS-Associat Rabbit Anti-SARS-Associat Mouse Anti-SARS-Associate Mouse Anti-SARS-Associate Mouse Anti-SARS Nucleocap Anti-SARS Nucleocapsid Pr Rabbit Anti-SARS-CoV M Pr Cellufine Formyl , 50 ml Cellufine Formyl Media Cellufine Formyl , 500 ml Cellufine Formyl Media

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Construction and characterization of single-chain variable fragment antibody library derived from germline rearranged immunoglobulin variable genes.

Antibody repertoires for library construction are conventionally harvested from mRNAs of immune cells. To examine whether germline rearranged immunoglobulin (Ig) variable region genes could be used as source of antibody repertoire, an immunized phage-displayed scFv library was prepared using splenocytic genomic DNA as template. In addition, a novel frame-shifting PCR (fsPCR) step was introduced to rescue stop codon and to enhance diversity of the complementarity-determining region 3 (CDR3). The germline scFv library was initially characterized against the hapten antigen phenyloxazolone (phOx). Sequence analysis of the phOx-selective scFvs indicated that the CDRs consisted of novel as well as conserved motifs. In order to illustrate that the diversity of CDR3 was increased by the fsPCR step, a second scFv library was constructed using a single scFv clone L3G7C as a template. Despite showing similar binding characteristics towards phOx, the scFv clones that were obtained from the L3G7C-derived antibody library gave a lower non-specific binding than that of the parental L3G7C clone. To determine whether germline library represented the endogenous immune status, specific scFv clones for nucleocapsid (N) protein of SARS-associated coronavirus (SCoV) were obtained both from naïve and immunized germline scFv libraries. Both libraries yielded specific anti-N scFvs that exhibited similar binding characteristics towards recombinant N protein, except the immunized library gave a larger number of specific anti-N scFv, and clones with identical nucleotide sequences were found. In conclusion, highly diversified antibody library can be efficiently constructed using germline rearranged immunoglobulin variable genes as source of antibody repertoires and fsPCR to diversify the CDR3.

1214 related Products with: Construction and characterization of single-chain variable fragment antibody library derived from germline rearranged immunoglobulin variable genes.

Myosin Light Chain 2 (Pho Androgen Receptor (Phosph Androgen Receptor (Phosph Androgen Receptor (Ab 650 Myosin Light Chain 2 (Ab immunoglobulin superfamil thymic dendritic cell-der Beta Amyloid (40) ELISA K Beta Amyloid (1 42) ELISA Anti PDX1 Polyclonal Anti Rabbit Anti-Cytochrome b2 Proteins and Antibodies H

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SARS coronavirus nucleocapsid protein monoclonal antibodies developed using a prokaryotic expressed protein.

Immunological detection of viruses and their components using monoclonal antibodies (MAbs) is a powerful diagnostic method. Here we report a detailed method for the establishment of MAbs against severe acute respiratory syndrome coronavirus (SARS-CoV). To express and purify the nucleocapsid protein (N protein) of SARS-CoV and generate MAbs against the N protein, gene encoding N protein was separated into two parts according to the prediction of epitopes and cloned into pET32a(+), respectively. Expression of the target proteins were induced by M isopropyl-β-thio-D-galactopyranoside (IPTG) and purified by a single-step affinity chromatography on a Ni-NTA column. BALB/c mice were immunized with the purified recombinant proteins to prepare MAbs by hybridoma technique. The reactivity and specificity of the MAbs were analyzed by ELISA and Western blot analysis. Seven MAbs against N1 and two MAbs against N2 were obtained. In the present study, recombinant SARS-CoV N protein was expressed and purified and nine specific MAbs against SARS-CoV N protein were obtained successfully. This panel of anti-N MAbs may be used as a tool for rapid and specific diagnosis of SARS-CoV.

1717 related Products with: SARS coronavirus nucleocapsid protein monoclonal antibodies developed using a prokaryotic expressed protein.

Rabbit Anti-SARS Virus Nu Rabbit Anti-SARS-Associat Rabbit Anti-SARS-Associat Mouse Anti-SARS-Associate Mouse Anti-SARS-Associate Mouse Anti-SARS Nucleocap Anti-SARS Nucleocapsid Pr Rabbit Anti-SARS-Associat Rabbit Anti-SARS-Associat Rabbit Anti-SARS-Associat Rabbit Anti-SARS-Associat Anti-SARS Spike Protein I

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Development, characterization, and application of monoclonal antibodies against severe acute respiratory syndrome coronavirus nucleocapsid protein.

Five monoclonal antibodies (MAbs) against recombinant nucleocapsid protein (NP) of severe acute respiratory syndrome (SARS)-causing coronavirus (CoV) were developed by hybridoma technology. Epitope mapping by Western blotting showed that these anti-SARS-CoV NP MAbs bind to distinct domains of NP. These anti-SARS-CoV NP MAbs, with their high specificity, are potentially ideal candidates for developing early and sensitive diagnostic assays for SARS-CoV.

1519 related Products with: Development, characterization, and application of monoclonal antibodies against severe acute respiratory syndrome coronavirus nucleocapsid protein.

Rabbit Anti-SARS Virus Nu Rabbit Anti-SARS-Associat Rabbit Anti-SARS-Associat Mouse Anti-SARS-Associate Mouse Anti-SARS-Associate steroidogenic acute regul Mouse Anti-SARS Nucleocap Anti-SARS Nucleocapsid Pr Rabbit Anti-Rat Androgen Mouse monoclonal anti-fla Rat monoclonal anti mouse Rat monoclonal anti mouse

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Protein transfer enhances cellular immune responses to DNA vaccination against SARS-CoV.

The current DNA vaccine formulations are not optimal for stimulation of CD8(+) T cells, which are required for clearing virally-infected cells. Here we show that CD8(+) T cell-stimulating activity can be effectively augmented by combining DNA vaccination with protein transfer. C57BL/6 mice were injected intramuscularly with an anti-SARS-CoV DNA vaccine admixed with a lipid-derived conjugate of 4-1BBL, a potential CD8(+) T-cell co-stimulator. The inclusion of the lipidated co-stimulator greatly enhanced cellular immune responses, especially the CTL response, induced by the DNA vaccine. The adjuvant effect of 4-1BBL was lipidation-dependent, indicating that it functions as a cell membrane-anchored co-stimulator. Results of our study suggest, for the first time, that muscle cells may be modified in situ, at the DNA injection site, into APC-like cells to allow direct priming of CD8(+) T cells and thereby improve the efficacy of DNA vaccines.

2440 related Products with: Protein transfer enhances cellular immune responses to DNA vaccination against SARS-CoV.

Rabbit Anti-SARS-CoV M Pr Human Dnak (HSP70) His ta E. coli SSB (Single Stran E. coli SSB (Single Stran E. coli SSB (Single Stran E. coli SSB (Single Stran Taq SSB (Single Stranded Taq SSB (Single Stranded Rabbit Anti-SARS Virus Nu Rabbit Anti-SARS-Associat Rabbit Anti-SARS-Associat Mouse Anti-SARS-Associate

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Mouse studies of SARS coronavirus-specific immune responses to recombinant replication-defective adenovirus expressing SARS coronavirus N protein.

1. A recombinant adenovirus encoding SARS coronavirus(SARS-CoV) nucleocapsid protein (rAd-N) was constructed. 2. The ability of the rAd-N to induce anti-SARS-CoV N antibody production and cellular immune responses was evaluated in an HLA-A2.1/Kb transgenic mouse model.

1429 related Products with: Mouse studies of SARS coronavirus-specific immune responses to recombinant replication-defective adenovirus expressing SARS coronavirus N protein.

Mouse Anti-SARS-Associate Mouse Anti-SARS-Associate Recombinant Viral antige Recombinant Viral antige Rabbit Anti-SARS-Associat Rabbit Anti-SARS-Associat Recombinant Viral antige Recombinant Viral antige Recombinant Viral antige SARS Associated Coronavir Recombinant Viral antige SARS Associated Coronavir

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The expression and antigenicity of a truncated spike-nucleocapsid fusion protein of severe acute respiratory syndrome-associated coronavirus.

In the absence of effective drugs, controlling SARS relies on the rapid identification of cases and appropriate management of the close contacts, or effective vaccines for SARS. Therefore, developing specific and sensitive laboratory tests for SARS as well as effective vaccines are necessary for national authorities.

1473 related Products with: The expression and antigenicity of a truncated spike-nucleocapsid fusion protein of severe acute respiratory syndrome-associated coronavirus.

Rabbit Anti-SARS-Associat Rabbit Anti-SARS-Associat Mouse Anti-SARS-Associate Mouse Anti-SARS-Associate Rabbit Anti-SARS-Associat Rabbit Anti-SARS-Associat Rabbit Anti-SARS-Associat Recombinant Viral antige Recombinant Viral antige Recombinant Viral antige SARS Associated Coronavir Recombinant Viral antige

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Prior immunization with severe acute respiratory syndrome (SARS)-associated coronavirus (SARS-CoV) nucleocapsid protein causes severe pneumonia in mice infected with SARS-CoV.

The details of the mechanism by which severe acute respiratory syndrome-associated coronavirus (SARS-CoV) causes severe pneumonia are unclear. We investigated the immune responses and pathologies of SARS-CoV-infected BALB/c mice that were immunized intradermally with recombinant vaccinia virus (VV) that expressed either the SARS-CoV spike (S) protein (LC16m8rVV-S) or simultaneously all the structural proteins, including the nucleocapsid (N), membrane (M), envelope (E), and S proteins (LC16m8rVV-NMES) 7-8 wk before intranasal SARS-CoV infection. The LC16m8rVV-NMES-immunized group exhibited as severe pneumonia as the control groups, although LC16m8rVV-NMES significantly decreased the pulmonary SARS-CoV titer to the same extent as LC16m8rVV-S. To identify the cause of the exacerbated pneumonia, BALB/c mice were immunized with recombinant VV that expressed the individual structural proteins of SARS-CoV (LC16mOrVV-N, -M, -E, -S) with or without LC16mOrVV-S (i.e., LC16mOrVV-N, LC16mOrVV-M, LC16mOrVV-E, or LC16mOrVV-S alone or LC16mOrVV-N + LC16mOrVV-S, LC16mOrVV-M + LC16mOrVV-S, or LC16mOrVV-E + LC16mOrVV-S), and infected with SARS-CoV more than 4 wk later. Both LC16mOrVV-N-immunized mice and LC16mOrVV-N + LC16mOrVV-S-immunized mice exhibited severe pneumonia. Furthermore, LC16mOrVV-N-immunized mice upon infection exhibited significant up-regulation of both Th1 (IFN-gamma, IL-2) and Th2 (IL-4, IL-5) cytokines and down-regulation of anti-inflammatory cytokines (IL-10, TGF-beta), resulting in robust infiltration of neutrophils, eosinophils, and lymphocytes into the lung, as well as thickening of the alveolar epithelium. These results suggest that an excessive host immune response against the nucleocapsid protein of SARS-CoV is involved in severe pneumonia caused by SARS-CoV infection. These findings increase our understanding of the pathogenesis of SARS.

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Rabbit Anti-SARS-Associat Rabbit Anti-SARS-Associat Mouse Anti-SARS-Associate Mouse Anti-SARS-Associate Rabbit Anti-SARS-CoV M Pr Recombinant Viral antige Recombinant Viral antige Rabbit Anti-SARS Virus Nu Rabbit Anti-SARS-Associat Rabbit Anti-SARS-Associat Rabbit Anti-SARS-Associat Rabbit Anti-SARS-Associat

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Induction of neutralising antibodies and cellular immune responses against SARS coronavirus by recombinant measles viruses.

Live attenuated recombinant measles viruses (rMV) expressing a codon-optimised spike glycoprotein (S) or nucleocapsid protein (N) of severe acute respiratory syndrome-associated coronavirus (SARS-CoV) were generated (rMV-S and rMV-N). Both recombinant viruses stably expressed the corresponding SARS-CoV proteins, grew to similar end titres as the parental strain and induced high antibody titres against MV and the vectored SARS-CoV antigens (S and N) in transgenic mice susceptible to measles infection. The antibodies induced by rMV-S had a high neutralising effect on SARS-CoV as well as on MV. Moreover, significant N-specific cellular immune responses were measured by IFN-gamma ELISPOT assays. The pre-existence of anti-MV antibodies induced by the initial immunisation dose did not inhibit boost of anti-S and anti-N antibodies. Immunisations comprising a mixture of rMV-S and rMV-N induced immune responses similar in magnitude to that of vaccine components administered separately. These data support the suitability of MV as a bivalent candidate vaccine vector against MV and emerging viruses such as SARS-CoV.

2864 related Products with: Induction of neutralising antibodies and cellular immune responses against SARS coronavirus by recombinant measles viruses.

Recombinant Viral antige Recombinant Viral antige Recombinant Viral antige Recombinant Viral antige Recombinant Viral antige SARS Associated Coronavir Recombinant Viral antige SARS Associated Coronavir Recombinant Viral antige SARS Associated Coronavir Measles Virus Nucleoprote Measles Virus nucleoprote

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