Development of a Novel Recombinant Heamagglutinin-Neuramindase Elisa (rHN-ELISA) for Evaluation of Humoral Immunity in Chicken Vaccinated Against Newcastle Disease Virus (NDV)

Moon, Ho-Jin; Park, Jung Eun; Hyeon, Yoon; Cruz, Deu John M.; Kim, Chul-Joong; Shin, Hyun‐Jin

doi:10.3923/javaa.2010.2932.2939

Cited by 1 publication

(4 citation statements)

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“…A comparison of the endpoint titers showed that there are not statistically significant differences. These results were in line with the findings of other researchers using similar epitopic or the full-length of F or HN protein for immunogenicity in the animal models (16, 17, 19). Furthermore, the detection of the HN and F glycoproteins with the individually produced antibodies revealed the potential application of these antibodies as probes for the viral glycoprotein in immunological analyses.…”

Section: Discussionsupporting

confidence: 92%

“…In fact, recombinant proteins which expressed in the E. coli cytoplasm is partially insoluble due to hydrophobic residues which distributed on the surface of the proteins (33). Other studies have reported the soluble expression of HN or F in E. coli host in which the vectors with a fusion partner such as Trx and NusA have been used (16, 17). It was revealed that the expression of subunits of the heteromultimeric proteins in the soluble form sometimes results in their aggregation as the inclusion body in absence of a proper fusion tag (33).…”

Section: Discussionmentioning

confidence: 99%

“…Results of the experiment showed that the muti-epitopic recombinant HN and F antigens expressed in the E. coli could elicit a sufficient rise of antibody in the immunized mice for detection by ELISA as well as standard Newcastle vaccine which composed of the whole structural NDV proteins. On the other hand, the commercial strains that are currently being used for detection of NDV induce a high amount of antibodies which may not directly produce against HN or F antigens, because NDV structural proteins like nucleocapsid protein (NP) are also associated with the high immunogenicity properties, as well (17, 43).…”

Section: Discussionmentioning

confidence: 99%

“…However, there is relatively limited information about the immunological properties of the recombinant surface glycoproteins of the NDV expressed in the E. coli system. There are a few reports about the successful expression and purification of NDV glycoproteins in the E. coli in which the recombinant HN was used as main the component in the ELISA based method (17) and purification of the immunogenic F protein that were studied elsewhere (18, 19).…”

Section: Introductionmentioning

confidence: 99%

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Immunogenicity of the Multi-Epitopic Recombinant Glycoproteins of Newcastle Disease Virus: Implications for the Serodiagnosis Applications

et al. 2018

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Background Newcastle disease virus (NDV) is a dangerous viral disease, infecting a broad range of birds, and has a fatal effect on the poultry industries. The attachment and consequently fusion of the virus to the host cell membrane is directed by the two superficial glycoproteins, the hemagglutinin-neuraminidase (HN) and the fusion (F) which is considered as the important targets for the poultry immune response. Objectives The principal goal of this investigation was to realize the potential efficacy of the E. coli expression system for the production of the multi-epitopic HN, and F proteins with respect to the ability for the stimulation of the immune system and production of the cross-reactive antibodies in mice. Materials and Methods The recombinant HN and F (rHN, rF) have accumulated almost 40% of the total bacterial proteins. The presence of rHN and rF proteins recognized by the Western blotting with specific anti-HN, anti-F, anti-Newcastle B1, and anti-poly 6x His-tag antibodies. Furthermore, both rHN and rF have shown the specific reactivity against the Newcastle B1 antiserum as a standard strain. Results The ELISA analysis showed that the higher dilutions of the antibody against Newcastle B1 could react with the as least quantity as 100 ng of the purified rHN, and rF. Cross-reactivity analysis of the sera from the mice immunized with Newcastle B1 in two time points indicated that the raise of anti-Newcastle B1, anti-HN and anti-F antibodies peaked at 28 days post immunization (dpi). Moreover, temporal variation in IgG titration between both time points was significant at 5% probability level. Conclusion The results provided valuable information about the cross-reactivity patterns and biological activity of the multi-epitopic proteins compared to the NDV standard strain which was determined by the Western blotting and ELISA.

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Section: Discussionsupporting

confidence: 92%