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A single-domain antibody inhibits SFTSV and mitigates virus-induced pathogenesis in vivo
Xilin Wu, Yanlei Li, Bilian Huang, Xiaohua Ma, Linjing Zhu, Nan Zheng, Shijie Xu, Waqas Nawaz, Changping Xu, Zhiwei Wu
Xilin Wu, Yanlei Li, Bilian Huang, Xiaohua Ma, Linjing Zhu, Nan Zheng, Shijie Xu, Waqas Nawaz, Changping Xu, Zhiwei Wu
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Research Article Infectious disease Therapeutics

A single-domain antibody inhibits SFTSV and mitigates virus-induced pathogenesis in vivo

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Abstract

Severe fever with thrombocytopenia syndrome virus (SFTSV) is a novel tick-borne bunyavirus that recently emerged in East Asian countries. SFTS is characterized by high fever, thrombocytopenia, leukopenia, multiorgan failure, and hemorrhage with case fatality rates of 6.3% to 30%. Neither antivirals nor vaccines are available at present. We previously demonstrated that neutralizing antibodies specific for SFTSV glycoprotein (Gn) played a vital role in the survival of patients with SFTS. Nanobodies from camels present unique properties, such as thermostability, high affinity, and low immunogenicity. In the current study, mammalian expressed SFTSV Gn was used to immunize a camel, and functional nanobodies were isolated from the B cell nanobody library constructed from the immunized animal. Clone SNB02 was selected for in-depth analysis for its inhibition of SFTSV replication both in vitro and in vivo. We showed that SNB02 potently inhibited SFTSV infection and prevented thrombocytopenia in a humanized mouse model and is a potential candidate for therapeutics.

Authors

Xilin Wu, Yanlei Li, Bilian Huang, Xiaohua Ma, Linjing Zhu, Nan Zheng, Shijie Xu, Waqas Nawaz, Changping Xu, Zhiwei Wu

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Figure 2

The biopanning and screening of the sGn-VHH antibody phage library.

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The biopanning and screening of the sGn-VHH antibody phage library.
(A) ...
(A) The binding of the phage library with sGn via phage ELISA. Lib is the phage library of sGn VHH; 1st, 2nd, and 3rd are the phage library after panning on 1 round, 2 rounds, and 3 rounds of sGn enrichment, respectively. (B) Ninety-five single clones of phages from the sGn-VHH library after the second and third enrichment of sGN were analyzed by phage ELISA. (C) The summary of bacterial supernatant binding with sGn protein tested by phage ELISA. The y axis shows the ratio of the readout of sGn binding to the readout of blank binding at OD 450 nm. Each square represents bacterial supernatant from a single clone of 1399 clones. (D) Positive binding VHHs inhibiting SFTSV infection, where each dot represents 1 VHH clone supernatant. Dashed line is located at 50% relative inhibition.

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