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Characterization of chicken-derived antibody against Alpha-Enolase of Streptococcus pneumoniae
Chi-Hsin Lee1, Chao-Jung Wu1, Yi-Yuan Yang2
1School of Medical Laboratory Science and Biotechnology, College of Medical Science and Technology, Taipei Medical University, Taipei 110301, Taiwan; Ph.D. Program in Medical Biotechnology, College of Medical Science and Technology, Taipei Medical University, Taipei 110301, Taiwan.
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.
Insights
Researchers developed novel single-chain variable fragment (scFv) antibodies targeting Streptococcus pneumoniae alpha-enolase (spEno1). These antibodies show potential for immunotherapeutic strategies against pneumococcal infections.
Area of Science:
- Microbiology
- Immunology
- Biotechnology
Background:
- Streptococcus pneumoniae causes severe infections like pneumonia and meningitis.
- Antimicrobial resistance in S. pneumoniae is a growing global health concern.
- Targeting surface proteins like alpha-enolase (spEno1) offers alternative therapeutic strategies.
Purpose of the Study:
- To develop novel monoclonal antibodies against S. pneumoniae alpha-enolase (spEno1).
- To evaluate the efficacy of these antibodies in inhibiting bacterial virulence factors.
Main Methods:
- Recombinant spEno1 protein purification and chicken immunization for IgY antibody generation.
- Construction and screening of single-chain variable fragment (scFv) antibody libraries using phage display technology.
- Evaluation of antibody binding activity, epitope mapping, and inhibition assays.
Main Results:
- Ten scFv antibodies against spEno1 were successfully identified and characterized.
- These scFv antibodies demonstrated specific binding to spEno1.
- The antibodies effectively inhibited the interaction between spEno1 and human plasminogen.
Conclusions:
- The generated scFv antibodies show promise as a potential immunotherapeutic agent against S. pneumoniae.
- Targeting spEno1 represents a viable strategy to combat pneumococcal infections, especially in the context of rising antibiotic resistance.
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