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J Virol Methods ; 276: 113791, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31778678

RESUMO

Following the recommended use of the inactivated poliovirus vaccine from Sabin strains (sIPV) by the WHO, a D antigen-specific neutralizing monoclonal antibody (mAb) 1G10 that recognized the human poliovirus type 1 Sabin strain (PV-I Sabin) was produced for D-antigen potency evaluation on sIPV. Study of the mAb 1G10 showed that it recognized a discontinuous conformational epitope of PV-I Sabin antigen. To identify this epitope quickly, easily and cost-effectively, clues to the epitope's identity were first obtained by employing a novel mimotope strategy based on a phage display library and "in silico" prediction. Then, the conformation of the epitope region, including the amino acid sequence, neutralizing sites, and location of this epitope, was identified using several classic epitope-mapping methods such as synthesized peptides analysis, neutralization assay and site-directed mutagenesis. The mimotope strategy may offer some guidance for achieving epitope identification in a more feasible and effective way. This mAb could be used in an in-house or national and international standard IPV D-antigen potency ELISA kit in the future.


Assuntos
Anticorpos Monoclonais/imunologia , Mapeamento de Epitopos/métodos , Epitopos/imunologia , Poliovirus/imunologia , Sequência de Aminoácidos , Antígenos Virais/imunologia , Linhagem Celular , Biologia Computacional , Humanos , Biblioteca de Peptídeos , Vacina Antipólio Oral/análise
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