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Bacterial Inner-membrane Display for Screening a Library of Antibody Fragments
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PRODUCTION OF SINGLE CHAIN FRAGMENT VARIABLE (scFv) ANTIBODY AGAINST PARANOSEMA LOCUSTAE ALPHA/BETA-HYDROLASE AND

V V Dolgikh, A A Tsarev, I V Senderskiy

    Tsitologiia
    |September 7, 2018
    PubMed

    Insights

    Researchers developed novel single chain antibodies (scFv-fragments) to detect microsporidian alpha/beta-hydrolase secretion. This tool confirms the enzyme

    Area of Science:

    • Parasitology
    • Molecular Biology
    • Mycology

    Background:

    • Microsporidia are fungal-related intracellular parasites impacting host cell physiology.
    • Secretion of proteins by microsporidia is a key mechanism for host manipulation.
    • Previous studies indicated secretion of Paranosema locustae alpha/beta-hydrolase, but its heterologous expression lacked secretion.

    Purpose of the Study:

    • To develop a tool for detecting microsporidian protein secretion.
    • To confirm the secretion of Paranosema locustae alpha/beta-hydrolase in infected locusts.
    • To investigate the role of secreted microsporidian hydrolases in host-parasite interactions.

    Main Methods:

    • Construction of a recombinant single chain antibody (scFv) library against infected locust fat body proteins.
    • Phage display technology for isolating scFv-fragments specific to P. locustae alpha/beta-hydrolase.
    • Immunoblotting and immunofluorescent microscopy using selected scFv-fragments for detection.

    Main Results:

    • Isolation of a specific mini-antibody (scFv-fragment) recognizing P. locustae alpha/beta-hydrolase.
    • Confirmation of the secretion of two distinct molecular weight forms of the hydrolase into infected host cells.
    • Demonstration of the utility of scFv-fragments in studying microsporidian secreted proteins.

    Conclusions:

    • Single chain antibodies are effective tools for studying microsporidian secreted proteins.
    • P. locustae alpha/beta-hydrolase is secreted in multiple forms, suggesting complex post-translational modifications or isoforms.
    • Further research using these tools can elucidate the function of microsporidian hydrolases in host-parasite relationships.

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