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Structural characterization of humanized nanobodies with neutralizing activity against the bordetella pertussis CyaA-hemolysin: Implications for a potential epitope of toxin-protective antigen
Malik A.A. - ไม่ระบุหน่วยงาน
ชื่อเรื่อง (EN): Structural characterization of humanized nanobodies with neutralizing activity against the bordetella pertussis CyaA-hemolysin: Implications for a potential epitope of toxin-protective antigen
ผู้แต่ง / หัวหน้าโครงการ (EN): Malik A.A.
บทคัดย่อ (EN): Previously, the 126-kDa CyaA-hemolysin (CyaA-Hly) fragment cloned from Bordetella pertussis—the causative agent of whooping cough—and functionally expressed in Escherichia coli was revealed as a key determinant for CyaA-mediated hemolysis against target erythrocytes. Here, phagemid-transfected E. coli clones producing nanobodies capable of binding to CyaA-Hly were selected from a humanized-camel VH/V H H phage-display library. Subsequently verified for binding activities by indirect ELISA and Western blotting, four CyaA-Hly-specific nanobodies were obtained and designated according to the presence/absence of V H H-hallmark amino acids as V H H2, VH5, VH18 and V H H37. In vitro neutralization assay revealed that all four ~17-kDa His-tagged VH/VHH nanobodies, in particular V H H37, which were over-expressed as inclusions and successfully unfolded-refolded, were able to effectively inhibit CyaA-Hly-mediated hemolysis. Phage-mimotope searching revealed that only peptides with sequence homologous to Linker 1 connecting Blocks I and II within the CyaA-RTX subdomain were able to bind to these four CyaA-Hly-specific nanobodies. Structural analysis of VHH37 via homology modeling and intermolecular docking confirmed that this humanized nanobody directly interacts with CyaA-RTX/Linker 1 through multiple hydrogen and ionic bonds. Altogether, our present data demonstrate that CyaA-RTX/Linker 1 could serve as a potential epitope of CyaA-protective antigen that may be useful for development of peptide-based pertussis vaccines. Additionally, such toxin-specific nanobodies have a potential for test-driven development of a ready-to-use therapeutic in passive immunization for mitigation of disease severity. © 2016 by the authors; licensee MDPI, Basel, Switzerland.
บทคัดย่อ: ไม่พบข้อมูลจากหน่วยงานต้นทาง
ภาษา (EN): en
เอกสารแนบ (EN): https://www.scopus.com/inward/record.uri?eid=2-s2.0-84976328152&doi=10.3390%2ftoxins8040099&partnerID=40&md5=9f8b84226fc53c1a124e70f6cb1bc705
เผยแพร่โดย (EN): มหาวิทยาลัยมหิดล
คำสำคัญ (EN): Single-Domain Antibodies
เจ้าของลิขสิทธิ์ (EN): มหาวิทยาลัยมหิดล
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Malik A.A.. "Structural characterization of humanized nanobodies with neutralizing activity against the bordetella pertussis CyaA-hemolysin: Implications for a potential epitope of toxin-protective antigen". ไม่ระบุหน่วยงาน. ม.ป.ป.

Malik A.A.. "Structural characterization of humanized nanobodies with neutralizing activity against the bordetella pertussis CyaA-hemolysin: Implications for a potential epitope of toxin-protective antigen". ไม่ระบุหน่วยงาน. ม.ป.ป.. Print.



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Structural characterization of humanized nanobodies with neutralizing activity against the bordetella pertussis CyaA-hemolysin: Implications for a potential epitope of toxin-protective antigen
Malik A.A.
มหาวิทยาลัยมหิดล
ไม่ระบุวันที่เผยแพร่
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