Characterization of chicken-derived antibody against Alpha-Enolase of Streptococcus pneumoniae

Chi Hsin Lee, Chao Jung Wu, Yi Yuan Yang, Wei Chu Wang, Sy Jye Leu, Cheng Tsang Wu, Pei Shih Kao, Ko Jiunn Liu, Bor Yu Tsai, Yu Wei Chiang, Yan Chiao Mao, Nhlanhla Benedict Dlamini, Jungshan Chang

Research output: Contribution to journalArticlepeer-review

Abstract

Streptococcus pneumoniae is a clinically relevant pathogen notorious for causing pneumonia, meningitis, and otitis media in immunocompromised patients. Currently, antibiotic therapy is the most efficient treatment for fighting pneumococcal infections. However, an arise in antimicrobial resistance in S. pneumoniae has become a serious health issue globally. To resolve the problem, alternative and cost-effective strategies, such as monoclonal antibody-based targeted therapy, are needed for combating bacterial infection. S. pneumoniae alpha-enolase (spEno1), which is thought to be a great target, is a surface protein that binds and converts human plasminogen to plasmin, leading to accelerated bacterial infections. We first purified recombinant spEno1 protein for chicken immunization to generate specific IgY antibodies. We next constructed two single-chain variable fragments (scFv) antibody libraries by phage display technology, containing 7.2 × 107 and 4.8 × 107 transformants. After bio-panning, ten scFv antibodies were obtained, and their binding activities to spEno1 were evaluated on ELISA, Western blot and IFA. The epitopes of spEno1 were identified by these scFv antibodies, which binding affinities were determined by competitive ELISA. Moreover, inhibition assay displayed that the scFv antibodies effectively inhibit the binding between spEno1 and human plasminogen. Overall, the results suggested that these scFv antibodies have the potential to serve as an immunotherapeutic drug against S. pneumoniae infections.

Original languageEnglish
Article number111476
JournalInternational Immunopharmacology
Volume128
DOIs
Publication statusPublished - Feb 15 2024

Keywords

  • Alpha-enolase (Eno1)
  • IgY
  • Phage display
  • S. pneumoniae
  • Single chain antibody (scFv)

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology
  • Pharmacology

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