A simple nanobody-based competitive ELISA to detect antibodies against African swine fever virus

Zhao, Jian; Zhu, Jiahong; Wang, Ying; Yang, Mengting; Zhang, Qiang; Zhang, Chong; Nan, Yuchen; Zhou, En‐Min; Sun, Yani; Zhao, Qin

doi:10.1016/j.virs.2022.09.004

Cited by 16 publications

(5 citation statements)

References 59 publications

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“…In comparison to indirect ELISA, the main advantage of blocking (competitive) ELISA is its high sensitivity to compositional differences in complex antigen mixtures, even when the specific detecting antibody is present in relatively small amounts (125). Previous research has demonstrated the successful implementation of blocking (competitive) ELISA techniques based on ASFV structural proteins, including p72 (87), p54 (88), p30 (89)(90)(91)(92) and non-structural protein pk205R (93). Wang M (94) and Wang et al (95) have developed a double-antigen sandwich ELISA utilizing specific enzyme-conjugated antigens, namely p30 protein and p72 protein, exhibiting a sensitivity range of 1:1280 to 1:12,800 (Table 3).…”

Section: Asfv Associated Antibodies Detection Elisamentioning

confidence: 99%

Current detection methods of African swine fever virus

Hu,

Tian,

Lai

et al. 2023

Front. Vet. Sci.

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African swine fever (ASF), caused by the African swine fever virus (ASFV), is a highly contagious and notifiable animal disease in domestic pigs and wild boars, as designated by the World Organization for Animal Health (WOAH). The effective diagnosis of ASF holds great importance in promptly controlling its spread due to its increasing prevalence and the continuous emergence of variant strains. This paper offers a comprehensive review of the most common and up-to-date methods established for various genes/proteins associated with ASFV. The discussed methods primarily focus on the detection of viral genomes or particles, as well as the detection of ASFV associated antibodies. It is anticipated that this paper will serve as a reference for choosing appropriate diagnostic methods in diverse application scenarios, while also provide direction for the development of innovative technologies in the future.

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Section: Asfv Associated Antibodies Detection Elisamentioning

confidence: 99%

Current detection methods of African swine fever virus

Hu,

Tian,

Lai

et al. 2023

Front. Vet. Sci.

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show abstract

“…Three replicates of each sample were run, and each experiment was independently repeated three times. The mean OD 450 values presented in Table S2, S3, S4, S5, S6, S7, S8 and S9 were calculated to obtain P/N values according to previous studies [44][45][46].…”

Section: Optimization Of Elisamentioning

confidence: 99%

Development of an enzyme-linked immunosorbent assay based on viral antigen capture by anti-spike glycoprotein monoclonal antibody for detecting immunoglobulin A antibodies against porcine epidemic diarrhea virus in milk

Yang

et al. 2023

BMC Vet Res

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Background Porcine epidemic diarrhea (PED), caused by PED virus (PEDV), is a severe enteric disease burdening the global swine industry in recent years. Especially, the mortality of PED in neonatal piglets approaches 100%. Maternal antibodies in milk, particularly immunoglobulin A (IgA) antibodies, are of great importance for protection neonatal suckling piglets against PEDV infection as passive lactogenic immunity. Therefore, appropriate detection methods are required for detecting PEDV IgA antibodies in milk. In the current study, we prepared monoclonal antibodies (mAbs) against PEDV spike (S) glycoprotein. An enzyme-linked immunosorbent assay (ELISA) was subsequently developed based on PEDV antigen capture by a specific anti-S mAb. Results The developed ELISA showed high sensitivity (the maximum dilution of milk samples up to 1:1280) and repeatability (coefficient of variation values < 10%) in detecting PEDV IgA antibody positive and negative milk samples. More importantly, the developed ELISA showed a high coincidence rate with a commercial ELISA kit for PEDV IgA antibody detection in clinical milk samples. Conclusions The developed ELISA in the current study is applicable for PEDV IgA antibody detection in milk samples, which is beneficial for evaluating vaccination efficacies and neonate immune status against the virus.

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“…Among ASFV structural proteins, the p30 protein is encoded by the viral gene CP204L and is abundantly expressed in the early stages of infection ( 12 , 13 ). CP204L is a membrane-localized and highly antigenic protein with phosphorylation sites ( 14 , 15 ).…”

Section: Introductionmentioning

confidence: 99%

SNX32 is a host restriction factor that degrades African swine fever virus CP204L via the RAB1B-dependent autophagy pathway

Yang,

Li,

Zhang

et al. 2024

J Virol

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African swine fever virus (ASFV) causes a highly contagious and deadly disease in domestic pigs and European wild boars, posing a severe threat to the global pig industry. ASFV CP204L, a highly immunogenic protein, is produced during the early stages of ASFV infection. However, the impact of CP204L protein-interacting partners on the outcome of ASFV infection is poorly understood. To accomplish this, coimmunoprecipitation and mass spectrometry analysis were conducted in ASFV-infected porcine alveolar macrophages (PAMs). We have demonstrated that sorting nexin 32 (SNX32) is a CP204L-binding protein and that CP204L interacted and colocalized with SNX32 in ASFV-infected PAMs. ASFV growth and replication were promoted by silencing SNX32 and suppressed by overexpressing SNX32. SNX32 degraded CP204L by recruiting the autophagy-related protein Ras-related protein Rab-1b (RAB1B). RAB1B overexpression inhibited ASFV replication, while knockdown of RAB1B had the opposite effect. Additionally, RAB1B, SNX32, and CP204L formed a complex upon ASFV infection. Taken together, this study demonstrates that SNX32 antagonizes ASFV growth and replication by recruiting the autophagy-related protein RAB1B. This finding extends our understanding of the interaction between ASFV CP204L and its host and provides new insights into exploring the relationship between ASFV infection and autophagy. IMPORTANCE African swine fever (ASF) is a highly contagious and acute hemorrhagic viral disease with a high mortality near 100% in domestic pigs. ASF virus (ASFV), which is the only member of the family Asfarviridae , is a dsDNA virus of great complexity and size, encoding more than 150 proteins. Currently, there are no available vaccines against ASFV. ASFV CP204L represents the most abundantly expressed viral protein early in infection and plays an important role in regulating ASFV replication. However, the mechanism by which the interaction between ASFV CP204L and host proteins affects ASFV replication remains unclear. In this study, we demonstrated that the cellular protein SNX32 interacted with CP204L and degraded CP204L by upregulating the autophagy-related protein RAB1B. In summary, this study will help us understand the interaction mechanism between CP204L and its host upon infection and provide new insights for the development of vaccines and antiviral drugs.

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A simple nanobody-based competitive ELISA to detect antibodies against African swine fever virus

Cited by 16 publications

References 59 publications

Current detection methods of African swine fever virus

Current detection methods of African swine fever virus

Development of an enzyme-linked immunosorbent assay based on viral antigen capture by anti-spike glycoprotein monoclonal antibody for detecting immunoglobulin A antibodies against porcine epidemic diarrhea virus in milk

SNX32 is a host restriction factor that degrades African swine fever virus CP204L via the RAB1B-dependent autophagy pathway

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