Screening and characterization of inhibitory vNAR targeting nanodisc-assembled influenza M2 proteins

Chuandi Yu; Wen Ding; Lei Zhu; Yuhang Zhou; Yingkui Dong; Ling Li; Juanjuan Liu; Yizhuo Wang; Zehua Li; Lina Zhu; FaJun Chen; Maosen Ruan; Dongming Qian; Yujuan Wang; Bo Wu; Huangtao Xu; Ming Li; Yunchen Bi; Hao Wang; Weiqian Wang; Peng Chao; Lei Xing; Bing Shen; Han Dai; Lisha Zha; Hongxin Zhao; Junfeng Wang

doi:10.1016/j.isci.2022.105736

Screening and characterization of inhibitory vNAR targeting nanodisc-assembled influenza M2 proteins

iScience. 2022 Dec 5;26(1):105736. doi: 10.1016/j.isci.2022.105736. eCollection 2023 Jan 20.

Authors

Chuandi Yu^{1

2}, Wen Ding³, Lei Zhu¹, Yuhang Zhou⁴, Yingkui Dong^{1

5}, Ling Li¹, Juanjuan Liu¹, Yizhuo Wang^{1

2}, Zehua Li^{1

2}, Lina Zhu^{1

2}, FaJun Chen^{1

3}, Maosen Ruan¹, Dongming Qian⁶, Yujuan Wang¹, Bo Wu¹, Huangtao Xu¹, Ming Li^{1

2}, Yunchen Bi^{7

8}, Hao Wang^{7

8}, Weiqian Wang⁹, Peng Chao⁹, Lei Xing^{1

2}, Bing Shen³, Han Dai¹, Lisha Zha⁴, Hongxin Zhao^{1

6}, Junfeng Wang^{1

2

5}

Affiliations

¹ High Magnetic Field Laboratory, Key Laboratory of High Magnetic Field and Ion Beam Physical Biology, Hefei Institutes of Physical Science, Chinese Academy of Sciences, Hefei, Anhui 230031, China.
² University of Science and Technology of China, Hefei, Anhui 230026, China.
³ School of Basic Medical Sciences, Anhui Medical University, Hefei, Anhui 230032, China.
⁴ International Immunology Centre, Anhui Agricultural University, Hefei, Anhui 230036, China.
⁵ Institute of Physical Science and Information Technology, Anhui University, Hefei, Anhui 230039, China.
⁶ Hefei China Science Longwood Biological Technology Co., Ltd. Hefei, Anhui 230088, China.
⁷ CAS and Shandong Province Key Laboratory of Experimental Marine Biology, Center for OceanMega-Science, Institute of Oceanology, Chinese Academy of Sciences, Qingdao, Shandong 266071, China.
⁸ Marine Biology and Biotechnology Laboratory, Pilot National Laboratory for Marine Science and Technology (Qingdao), Qingdao, Shandong 266237, China.
⁹ National Facility for Protein Science in Shanghai, Shanghai Advanced Research Institute, Chinese Academy of Science, Shanghai 201210, China.

Abstract

Influenza A virus poses a constant challenge to human health. The highly conserved influenza matrix-2 (M2) protein is an attractive target for the development of a universal antibody-based drug. However, screening using antigens with subphysiological conformation in a nonmembrane environment significantly reduces the generation of efficient antibodies. Here, M2(1-46) was incorporated into nanodiscs (M2-nanodiscs) with M2 in a membrane-embedded tetrameric conformation, closely resembling its natural physiological state in the influenza viral envelope. M2-nanodisc generation, an antigen, was followed by Chiloscyllium plagiosum immunization. The functional vNARs were selected by phage display panning strategy from the shark immune library. One of the isolated vNARs, AM2H10, could specifically bind to tetrameric M2 instead of monomeric M2e (the ectodomain of M2 protein). Furthermore, AM2H10 blocked ion influx through amantadine-sensitive and resistant M2 channels. Our findings indicated the possibility of developing functional shark nanobodies against various influenza viruses by targeting the M2 protein.

Keywords: Nanoparticles; Virology.