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Production and characterization of single-chain variable fragment antibodies targeting the breast cancer tumor marker nectin-4.
Liu, Ching-Hsuan; Leu, Sy-Jye; Lee, Chi-Hsin; Lin, Cheng-Yuan; Wang, Wei-Chu; Tsai, Bor-Yu; Lee, Yu-Ching; Chen, Chi-Long; Yang, Yi-Yuan; Lin, Liang-Tzung.
Afiliación
  • Liu CH; Department of Microbiology and Immunology, School of Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan.
  • Leu SJ; Department of Microbiology & Immunology, Dalhousie University, Halifax, Nova Scotia, Canada.
  • Lee CH; Department of Microbiology and Immunology, School of Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan.
  • Lin CY; Graduate Institute of Medical Sciences, College of Medicine, Taipei Medical University, Taipei, Taiwan.
  • Wang WC; School of Medical Laboratory Science and Biotechnology, College of Medical Science and Technology, Taipei Medical University, Taipei, Taiwan.
  • Tsai BY; Ph.D. Program in Medical Biotechnology, College of Medical Science and Technology, Taipei Medical University, Taipei, Taiwan.
  • Lee YC; School of Medical Laboratory Science and Biotechnology, College of Medical Science and Technology, Taipei Medical University, Taipei, Taiwan.
  • Chen CL; Ph.D. Program in Medical Biotechnology, College of Medical Science and Technology, Taipei Medical University, Taipei, Taiwan.
  • Yang YY; Navi Bio-Therapeutics Inc., Taipei, Taiwan.
  • Lin LT; The Center of Translational Medicine, Taipei Medical University, Taipei, Taiwan.
Front Immunol ; 14: 1292019, 2023.
Article en En | MEDLINE | ID: mdl-38288120
ABSTRACT

Background:

Nectin-4 is a novel biomarker overexpressed in various types of cancer, including breast cancer, in which it has been associated with poor prognosis. Current literature suggests that nectin-4 has a role in cancer progression and may have prognostic and therapeutic implications. The present study aims to produce nectin-4-specific single-chain variable fragment (scFv) antibodies and evaluate their applications in breast cancer cell lines and clinical specimens.

Methods:

We generated recombinant nectin-4 ectodomain fragments as immunogens to immunize chickens and the chickens' immunoglobulin genes were amplified for construction of anti-nectin-4 scFv libraries using phage display. The binding capacities of the selected clones were evaluated with the recombinant nectin-4 fragments, breast cancer cell lines, and paraffin-embedded tissue sections using various laboratory approaches. The binding affinity and in silico docking profile were also characterized.

Results:

We have selected two clones (S21 and L4) from the libraries with superior binding capacity. S21 yielded higher signals when used as the primry antibody for western blot analysis and flow cytometry, whereas clone L4 generated cleaner and stronger signals in immunofluorescence and immunohistochemistry staining. In addition, both scFvs could diminish attachment-free cell aggregation of nectin-4-positive breast cancer cells. As results from ELISA indicated that L4 bound more efficiently to fixed nectin-4 ectodomain, molecular docking analysis was further performed and demonstrated that L4 possesses multiple polar contacts with nectin-4 and diversity in interacting residues.

Conclusion:

Overall, the nectin-4-specific scFvs could recognize nectin-4 expressed by breast cancer cells and have the merit of being further explored for potential diagnostic and therapeutic applications.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Anticuerpos de Cadena Única / Neoplasias Tipo de estudio: Prognostic_studies Límite: Animals Idioma: En Revista: Front Immunol Año: 2023 Tipo del documento: Article País de afiliación: Taiwán Pais de publicación: Suiza
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Anticuerpos de Cadena Única / Neoplasias Tipo de estudio: Prognostic_studies Límite: Animals Idioma: En Revista: Front Immunol Año: 2023 Tipo del documento: Article País de afiliación: Taiwán Pais de publicación: Suiza