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Baculovirus display of functional antibody Fab fragments

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Abstract

The generation of a recombinant baculovirus that displays antibody Fab fragments on the surface was investigated. A recombinant baculovirus was engineered so that the heavy chain (Hc; Fd fragment) of a mouse Fab fragment was expressed as a fusion to the N-terminus of baculovirus gp64, while the light chain of the Fab fragment was simultaneously expressed as a secretory protein. Following infection of Sf9 insect cells with the recombinant baculovirus, the culture supernatant was analyzed by enzyme-linked immunosorbent assay using antigen-coated microplates and either an anti-mouse IgG or an anti-gp64 antibody. A relatively strong signal was obtained in each case, showing antigen-binding activity in the culture supernatant. In western blot analysis of the culture supernatant using the anti-gp64 antibody, specific protein bands were detected at an electrophoretic mobility that coincided with the molecular weight of the Hc-gp64 fusion protein as well as that of gp64. Flow cytometry using a fluorescein isothiocyanate-conjugated antibody specific to mouse IgG successfully detected the Fab fragments on the surface of the Sf9 cells. These results suggest that immunologically functional antibody Fab fragments can be displayed on the surface of baculovirus particles, and that a fluorescence-activated cell sorter with a fluorescence-labeled antigen can isolate baculoviruses displaying specific Fab fragments. This successful baculovirus display of antibody Fab fragments may offer a novel approach for the efficient selection of specific antibodies.

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Acknowledgments

The authors thank Professor I. Fujii of the Department of Biological Science, Graduate School of Science, Osaka Prefecture University, for providing the plasmid pARA7-6D9Fab and the conjugate of hapten and BSA.

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Authors and Affiliations

  1. Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodai, Nada, Kobe, 657-8501, Japan

    Shinya Takada, Takafumi Ogawa, Kazusa Matsui, Tasuku Suzuki, Tomohisa Katsuda & Hideki Yamaji

  2. Institute of Pathology, Kyodo Byori, Inc., 2-7-12 Otsuwa, Nishi-ku, Kobe, 651-2112, Japan

    Takafumi Ogawa

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  1. Shinya Takada
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  2. Takafumi Ogawa
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  3. Kazusa Matsui
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  4. Tasuku Suzuki
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  5. Tomohisa Katsuda
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  6. Hideki Yamaji
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Correspondence to Hideki Yamaji.

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Takada, S., Ogawa, T., Matsui, K. et al. Baculovirus display of functional antibody Fab fragments. Cytotechnology 67, 741–747 (2015). https://doi.org/10.1007/s10616-015-9876-7

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  • DOI: https://doi.org/10.1007/s10616-015-9876-7

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