Computational Identification of Antibody Epitopes on the Dengue Virus NS1 Protein

Jones, Martina L.; Legge, Fiona S.; Lebani, Kebaneilwe; Mahler, Stephen M.; Young, Paul R.; Watterson, Daniel; Treutlein, Herbert; Zeng, Jun

doi:10.3390/molecules22040607

Cited by 13 publications

(19 citation statements)

References 41 publications

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“…This sequence was used as query for homology modelling. PDB:1QOK [22] structure showed highest percentage similarity to the 3IXY scFv antibody so it was selected as template to create scFv structure by homology modelling. Homology modelling of scFv sequence resulted in three models out of which the model with best DOPE score and PDF energy was selected for further analysis ( Fig 2 ).Dengue envelope protein (PDB:1OAN) was selected [21].…”

Section: Resultsmentioning

confidence: 99%

“…Recently, structure-guided computational approaches have been utilized for several applications in the field of antibody engineering for examples, mapping of neutralizing epitope [22] and antigen-antibody interaction analysis for identification of interacting CDR residues [23]. Such analysis could be helpful in reducing the extent of work required to develop an efficient and neutralizing antibody.…”

Section: Discussionmentioning

confidence: 99%

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Designing antibody against highly conserved region of dengue envelope protein by in silico screening of scFv mutant library

2019

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Dengue being one of the deadliest diseases of tropical regions, enforces to put continuous efforts for the development of vaccine and effective therapeutics. Most of the antibodies generated during dengue infection are non-neutralizing and cause antibody dependent enhancement. Hence, making a potent neutralizing antibody against all four dengue serotypes could be very effective for the treatment. However, designing a single antibody for all serotypes is difficult due to variation in protein sequences. Therefore, the objective is to identify conserved region of dengue envelope protein and then develop an antibody against that conserved region. Before advancing to the development of such an antibody, it is desirable to validate the interactions between antibody and dengue envelope protein. In silico analysis of such interactions provides a good platform to find out a suitable region to design and construct an antibody against it by analyzing antigen-antibody interaction before synthesizing the antibody. In this study, two highly conserved regions of dengue envelope protein were identified and an scFv was constructed against it. Both scFv and FuBc proteins were expressed in bacterial expression system and binding efficiency was analyzed by SPR analysis with KD value 2.3 μM. In order to improve binding efficiency, an in silico scFv mutant library was created which was virtually screened for higher binding efficiency. Six mutants with high binding efficiency were selected for further analysis. The binding ability of these mutants were predicted using simulation analysis which shows these mutations were stabilizing scFv-FuBc complex.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Designing antibody against highly conserved region of dengue envelope protein by in silico screening of scFv mutant library

2019

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“…Epitope-mapping studies have identified the sequence 193 AVHADMGYWIESALNDT 209 as the conformational epitope buried within a β -sheet structure of the dengue NS1 protein (16). The presence therein of net three negative charges conferred by the aspartic and glutamic acid residues may result in more stable immunocomplexes at higher temperature, as previously measured in other systems (17).…”

Section: Discussionmentioning

confidence: 78%

Febrile temperatures increase in vitro antibody affinity for malaria and dengue antigens

Stan

Françoso

Alves

et al. 2018

Preprint

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Fever is a regulated elevation in the body setpoint temperature and may arise as a result of infectious and noninfectious causes. While beneficial in modulating immune responses to infection, the potential of febrile temperatures in regulating antigen binding affinity to antibodies has not been explored. We have investigated this process under in vitro conditions using selected malaria or dengue antigens and specific monoclonal antibodies, and observed a marked increase in the affinity of these antibody-antigen complexes at 40˚C, compared to physiological (37˚C) or pathophysiological temperatures (42˚C). Induced thermal equilibration of the protein partners at these temperatures, prior to measurements, further increased their binding affinity. These results may indicate an unexpected beneficial and adaptive role for fever in vivo, and highlight the positive role of thermal priming in enhancing protein-protein affinity for samples of scarce availability.

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“…Epitope-mapping studies have identified the sequence 193 AVHADMGYWIESALNDT 209 as the conformational epitope from a β-sheet structure of the dengue NS1 protein [22]. The presence therein of three negative charges conferred by the aspartic and glutamic acid residues may result in more stable immune complexes at higher temperature, as previously measured in other systems [23].…”

Section: Resultsmentioning

confidence: 97%

Febrile temperatures increase in vitro antibody affinity for malarial and dengue antigens

Stan¹,

Françoso²,

Alves³

et al. 2019

PLoS Negl Trop Dis

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Fever is a regulated increase of the body temperature resulting from both infectious and non-infectious causes. Fever is known to play a role in modulating immune responses to infection, but the potential of febrile temperatures in regulating antigen binding affinity to antibodies has not been explored. Here we investigated this process under in vitro conditions using Isothermal titration calorimetry and ELISA. We used selected malarial and dengue antigens against specific monoclonal antibodies, and observed a marked increase in the affinity of these antibody-antigen complexes at 40°C, compared to physiological (37°C) or pathophysiological temperatures (42°C). Induced thermal equilibration of the protein partners at these temperatures in vitro , prior to measurements, further increased their binding affinity. These results suggest another positive and adaptive role for fever in vivo , and highlight the favourable role of thermal priming in enhancing protein-protein affinity for samples with limited availability.

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Computational Identification of Antibody Epitopes on the Dengue Virus NS1 Protein

Cited by 13 publications

References 41 publications

Designing antibody against highly conserved region of dengue envelope protein by in silico screening of scFv mutant library

Designing antibody against highly conserved region of dengue envelope protein by in silico screening of scFv mutant library

Febrile temperatures increase in vitro antibody affinity for malaria and dengue antigens

Febrile temperatures increase in vitro antibody affinity for malarial and dengue antigens

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