TY - JOUR
T1 - Characterization of chicken-derived antibody against Alpha-Enolase of Streptococcus pneumoniae
AU - Lee, Chi Hsin
AU - Wu, Chao Jung
AU - Yang, Yi Yuan
AU - Wang, Wei Chu
AU - Leu, Sy Jye
AU - Wu, Cheng Tsang
AU - Kao, Pei Shih
AU - Liu, Ko Jiunn
AU - Tsai, Bor Yu
AU - Chiang, Yu Wei
AU - Mao, Yan Chiao
AU - Benedict Dlamini, Nhlanhla
AU - Chang, Jungshan
N1 - Publisher Copyright:
© 2023
PY - 2024/2/15
Y1 - 2024/2/15
N2 - 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.
AB - 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.
KW - Alpha-enolase (Eno1)
KW - IgY
KW - Phage display
KW - S. pneumoniae
KW - Single chain antibody (scFv)
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U2 - 10.1016/j.intimp.2023.111476
DO - 10.1016/j.intimp.2023.111476
M3 - Article
C2 - 38185035
AN - SCOPUS:85181811612
SN - 1567-5769
VL - 128
JO - International Immunopharmacology
JF - International Immunopharmacology
M1 - 111476
ER -