Construction of a T7 phage display nanobody library for bio-panning and identification of chicken dendritic cell-specific binding nanobodies

文献类型： 外文期刊

第一作者： Xu, Hai

作者： Xu, Hai;Li, Ling;Hong, Weiming;Li, Ruiting;Guo, Zijie;Xu, Hai;Govinden, Roshini;Chenia, Hafizah Y.;Xu, Hai;Deng, Bihua;Hou, Jibo;Deng, Bihua

作者机构：

期刊名称：SCIENTIFIC REPORTS （ 影响因子：4.996； 五年影响因子：5.516 ）

ISSN： 2045-2322

年卷期： 2022 年 12 卷 1 期

页码：

收录情况： SCI

摘要： Dendritic cells (DCs) are the antigen-presenting cells that initiate and direct adaptive immune responses, and thus are critically important in vaccine design. Although DC-targeting vaccines have attracted attention, relevant studies on chicken are rare. A high diversity T7 phage display nanobody library was constructed for bio-panning of intact chicken bone marrow DCs to find DC-specific binding nanobodies. After three rounds of screening, 46 unique sequence phage clones were identified from 125 randomly selected phage clones. Several DC-binding phage clones were selected using the specificity assay. Phage-54, -74, -16 and -121 bound not only with chicken DCs, but also with duck and goose DCs. In vitro, confocal microscopy observation demonstrated that phage-54 and phage-74 efficiently adsorbed onto DCs within 15 min compared to T7-wt. The pull-down assay, however, did not detect any of the previously reported proteins for chicken DCs that could have interacted with the nanobodies displayed on phage-54 and phage-74. Nonetheless, Specified pathogen-free chickens immunized with phage-54 and phage-74 displayed higher levels of anti-p10 antibody than the T7-wt, indicating enhanced antibody production by nanobody mediated-DC targeting. Therefore, this study identified two avian (chicken, duck and goose) DC-specific binding nanobodies, which may be used for the development of DC-targeting vaccines.

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