Characterization of monoclonal antibodies against waterfowl parvoviruses VP3 protein

Yin, Xiuchen; Zhang, Shumei; Gao, Youlan; Li, Jinzhe; Tan, Shanzhong; Liu, Hongyu; Wu, Xiaoying; Chen, Yuhuan; Liu, Ming; Zhang, Yun

doi:10.1186/1743-422x-9-288

Cited by 13 publications

(9 citation statements)

References 25 publications

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“…Canine adenovirus 2 was propagated and purified, then used to produce mAbs as previously described (Huang et al, 2007;Yin et al, 2012). Briefly, five 6-week-old female BALB/c mice were immunized with 60 µg purified CAdV-2 emulsified with complete Freund's adjuvant (Sigma-Aldrich).…”

Section: Preparation Of Monoclonal Antibodiesmentioning

confidence: 99%

Development of an Immunochromatographic Strip for Rapid Detection of Canine Adenovirus

Wang

Wen

et al. 2019

Front. Microbiol.

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Although canine adenovirus (CAdV) is highly prevalent in dogs, there is currently a lack of a quick diagnostic method. In this study, we developed a rapid immunochromatographic strip (ICS) assay using colloidal gold coupled to CAdV-2-specific monoclonal antibodies (mAbs). BALB/c mice were immunized with a purified CAdV-2 suspension, and four mAbs (belonging to two different epitopes) were generated and designated as 2C1, 7D7, 10D1, and 4G1. Western blot and protein spectral analysis indicated that the hexon protein of CAdV-2 recognized all four mAbs. The colloidal gold-coupled 7D7 and 2C1 mAbs were chosen for inclusion in the rapid ICS assay. The optimal concentrations of the coating antibody (2C1), the capture antibody (7D7), and the goat anti-mouse antibody were 1.0 mg/ml, 10 µg/ml, and 2.0 mg/ml, respectively. The limit of detection was approximately 2.0 × 10 2 tissue culture infective dose (TCID 50 )/ml. Other common canine viruses were tested to evaluate the specificity of the ICS, and positive results were observed for only CAdV-1 and CAdV-2. The ICS test was conducted on 360 samples to detect CAdV, and the results were compared with those of polymerase chain reaction (PCR) tests. The ICS test was found to be a sufficiently sensitive and specific detection method for the convenient and rapid detection of CAdV.

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Section: Preparation Of Monoclonal Antibodiesmentioning

confidence: 99%

Development of an Immunochromatographic Strip for Rapid Detection of Canine Adenovirus

Wang

Wen

et al. 2019

Front. Microbiol.

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“…The mice received two boosts of 100 µg of plasmid DNA in sterile PBS twice within a 2-week interval. mAbs were produced using techniques similar to that described previously 16 – 18 . The mice were euthanized by anesthesia and the spleens were removed using aseptic techniques.…”

Section: Methodsmentioning

confidence: 99%

Mapping a Type-specific Epitope by Monoclonal Antibody against VP3 Protein of Duck Hepatitis A Type 1 Virus

Zhang

Meng

et al. 2017

Sci Rep

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Duck hepatitis A subtype 1 virus (DHAV-1) infection causes high mortality in ducklings, resulting in significant losses to duck industries. VP3 is a structural protein of DHAV-1. However, B-cell epitopes on VP3 have not been investigated. To stimulate VP3 antibody response, eukaryotic expression plasmid pCI-neo-VP3 was constructed and used as DNA immunogen to prepare mAbs. Western blot showed that 25.5 kDa VP3 could be detected by mAbs in duck embryo fibroblast (DEF) cells transfected with pCI-neo-VP3. Immunofluorescence assay showed that mAbs could specifically bind to DEF cells infected with DHAV-1. DAPI staining indicated that VP3 localizes to the cytoplasm and nucleus of DHAV-1 infected DEF. With neutralizing mAb 3B7, minimal epitope PSNI was mapped. Sequence alignment indicated that 205PSNI208 is highly conserved among DHAV-1, but different from those of DHAV-2 and DHAV-3. Epitope peptide reacted specifically with DHAV-1-positive duck sera by dot blotting, revealing PSNI is DHAV-1 type-specific epitope and the importance of these amino acids in antibody-epitope binding reactivity. These findings provided useful information for understanding the antigenicity of VP3 and might be valuable in the development of epitope-based vaccine or diagnostic kit for DHAV-1 infection and provide insights for understanding the pathogenesis of DHAV-1.

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“…The mice spleens were collected aseptically by using euthanized anesthesia techniques. mAbs were produced using techniques similar to that described previously [ 33 , 34 , 35 ]. Splenocytes were fused with SP2/0-Ag14 myeloma cells.…”

Section: Methodsmentioning

confidence: 99%

Characterization of Monoclonal Antibodies against HA Protein of H1N1 Swine Influenza Virus and Protective Efficacy against H1 Viruses in Mice

Liu

Xue

et al. 2017

Viruses

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H1N1 swine influenza viruses (SIV) are prevalent in pigs globally, and occasionally emerge in humans, which raises concern about their pandemic threats. To stimulate hemagglutination (HA) of A/Swine/Guangdong/LM/2004 (H1N1) (SW/GD/04) antibody response, eukaryotic expression plasmid pCI-neo-HA was constructed and used as an immunogen to prepare monoclonal antibodies (mAbs). Five mAbs (designed 8C4, 8C6, 9D6, 8A4, and 8B1) against HA protein were obtained and characterized. Western blot showed that the 70 kDa HA protein could be detected by all mAbs in MDCK cells infected with SW/GD/04. Three mAbs—8C4, 8C6, and 9D6—have hemagglutination inhibition (HI) and neutralization test (NT) activities, and 8C6 induces the highest HI and NT titers. The protection efficacy of 8C6 was investigated in BALB/c mice challenged with homologous or heterologous strains of the H1 subtype SIV. The results indicate that mAb 8C6 protected the mice from viral infections, especially the homologous strain, which was clearly demonstrated by the body weight changes and reduction of viral load. Thus, our findings document for the first time that mAb 8C6 might be of potential therapeutic value for H1 subtype SIV infection.

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Characterization of monoclonal antibodies against waterfowl parvoviruses VP3 protein

Cited by 13 publications

References 25 publications

Development of an Immunochromatographic Strip for Rapid Detection of Canine Adenovirus

Development of an Immunochromatographic Strip for Rapid Detection of Canine Adenovirus

Mapping a Type-specific Epitope by Monoclonal Antibody against VP3 Protein of Duck Hepatitis A Type 1 Virus

Characterization of Monoclonal Antibodies against HA Protein of H1N1 Swine Influenza Virus and Protective Efficacy against H1 Viruses in Mice

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