Immuno-dominant dengue NS1 peptides as antigens for production of monoclonal antibodies

Munasinghe, Erandi; Athapaththu, Maheshi; Abeyewickreme, W.

doi:10.3389/fmolb.2022.935456

Cited by 3 publications

(3 citation statements)

References 45 publications

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“…In many peptide antibody development projects, synthetic peptides are commonly used via conjugation to a carrier protein or with chemical modifications for tag introduction 17 , 19 – 22 . The general concern with such approaches is the potential modification of the peptide structure and its orientation on the carrier protein which could affect its accessibility for binding 23 .…”

Section: Discussionmentioning

confidence: 99%

Subtractive panning for the isolation of monoclonal PEPITEM peptide antibody by phage display

Alassiri,

Lai,

Ch’ng

et al. 2023

Sci Rep

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Antibody phage display is a key tool for the development of monoclonal antibodies against various targets. However, the development of anti-peptide antibodies is a challenging process due to the small size of peptides for binding. This makes anchoring of peptides a preferred approach for panning experiments. A common approach is by using streptavidin as the anchor protein to present biotinylated peptides for panning. Here, we propose the use of recombinant expression of the target peptide and an immunogenic protein as a fusion for panning. The peptide inhibitor of trans-endothelial migration (PEPITEM) peptide sequence was fused to the Mycobacterium tuberculosis (Mtb) α-crystalline (AC) as an anchor protein. The panning process was carried out by subtractive selection of the antibody library against the AC protein first, followed by binding to the library to PEPITEM fused AC (PEPI-AC). A unique monoclonal scFv antibodies with good specificity were identified. In conclusion, the use of an alternative anchor protein to present the peptide sequence coupled with subtractive panning allows for the identification of unique monoclonal antibodies against a peptide target.

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Section: Discussionmentioning

confidence: 99%

Subtractive panning for the isolation of monoclonal PEPITEM peptide antibody by phage display

Alassiri,

Lai,

Ch’ng

et al. 2023

Sci Rep

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“…DENV-RNA and NS1 were also detected in urine and saliva samples [ 10 , 51 , 65 , 66 , 67 ] ( Table 2 ). A study in Helsinki tested 14 Finnish travelers with DF, examining the presence of DENV-RNA and NS1 in saliva, urine and serum concurrently, and found that DENV-RNA in urine samples can be detectable at a later stage than serum samples [ 10 ] (Schemes Ι).…”

Section: The Role Of Ns1 In Dengue Fever Diagnosismentioning

confidence: 99%

The Role of NS1 Protein in the Diagnosis of Flavivirus Infections

Fisher

Lustig

Sklan

et al. 2023

Viruses

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Nonstructural protein 1 (NS1) is a glycoprotein among the flavivirus genus. It is found in both membrane-associated and soluble secreted forms, has an essential role in viral replication, and modulates the host immune response. NS1 is secreted from infected cells within hours after viral infection, and thus immunodetection of NS1 can be used for early serum diagnosis of dengue fever infections instead of real-time (RT)-PCR. This method is fast, simple, and affordable, and its availability could provide an easy point-of-care testing solution for developing countries. Early studies show that detecting NS1 in cerebrospinal fluid (CSF) samples is possible and can improve the surveillance of patients with dengue-associated neurological diseases. NS1 can be detected postmortem in tissue specimens. It can also be identified using noninvasive methods in urine, saliva, and dried blood spots, extending the availability and effective detection period. Recently, an enzyme-linked immunosorbent assay (ELISA) assay for detecting antibodies directed against Zika virus NS1 has been developed and used for diagnosing Zika infection. This NS1-based assay was significantly more specific than envelope protein-based assays, suggesting that similar assays might be more specific for other flaviviruses as well. This review summarizes the knowledge on flaviviruses’ NS1’s potential role in antigen and antibody diagnosis.

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“…The genome of the dengue virus contains a single open reading frame that encodes a polyprotein sequence, which can form three structural proteins (C, prM, and E) and seven nonstructural proteins, including NS1, NS2A, NS2B, NS3, NS4A, NS4B, and NS5 (Perera, Kuhn, 2008). The NS1 protein of DENV is highly conserved across all four serotypes and also highly immunogenic, stimulating an immune response in infected individuals (Lindenbach, Rice, 1999;Munasinghe et al, 2022). NS1 can exist intracellularly, in the cytoplasm, or on the cell surface and induce immune responses that protect against Flavivirus infections.…”

Section: Introductionmentioning

confidence: 99%

Prediction of conservative epitopes in the NS1 sequences of all four dengue virus serotypes

Xuan Cuong,

Minh Thuong,

Nhu Binh

et al. 2023

Vietnam J. Biotechnol.

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The Dengue virus, transmitted by Aedes aegypti mosquitoes, causes dengue fever, a life-threatening illness. NS1 (Nonstructural Protein 1) has been widely used as a biomarker for detecting dengue due to its early occurrence, high sensitivity, and specificity. The main objective of this study was to design an epitope-rich peptide capable of identifying all four dengue serotypes using homology modeling. Homology protein modeling, a computational approach, predicts the 3D structure of a protein by comparing its sequence to a known template. To achieve this goal, various bioinformatics tools, such as Blast/NCBI, BepiPred, Discotope 2.0, and HADDOCK, were utilized. As a result, there were 18 consensus sequences for the four Dengue serotypes from amino acid 12 to 344. However, the eight of them have shown the most highly scoring conservative sequence segments in BepiBred, Antigenicity, Beta turn, Surface accessibility, Flexibility, and Hydrophilicity. In the results of predicting epitopes, three conservative segments predicted to be the best antigens were selected. Combining the resulting parts, two potential NS1 peptides, containing 149 amino acids from positions 112 to 260, were identified and characterized in all four DENV strains. It is the sequence segment that will be selected for creating materials such as recombinant antigens and monoclonal antibodies for the diagnosis of dengue hemorrhagic fever in the next study.

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Immuno-dominant dengue NS1 peptides as antigens for production of monoclonal antibodies

Cited by 3 publications

References 45 publications

Subtractive panning for the isolation of monoclonal PEPITEM peptide antibody by phage display

Subtractive panning for the isolation of monoclonal PEPITEM peptide antibody by phage display

The Role of NS1 Protein in the Diagnosis of Flavivirus Infections

Prediction of conservative epitopes in the NS1 sequences of all four dengue virus serotypes

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