https://scholars.lib.ntu.edu.tw/handle/123456789/626684
標題: | Tag-Free SARS-CoV-2 Receptor Binding Domain (RBD), but Not C-Terminal Tagged SARS-CoV-2 RBD, Induces a Rapid and Potent Neutralizing Antibody Response | 作者: | Lin, Ting-Wei Huang, Ping-Han Liao, Bo-Hung TAI-LING CHAO Tsai, Ya-Min SHIH-CHUNG CHANG SUI-YUAN CHANG HUI-WEN CHEN |
關鍵字: | COVID-19; Delta variant; His-tag RBD; neutralizing antibodies; tag-free RBD; vaccine | 公開日期: | 30-十月-2022 | 出版社: | MDPI | 卷: | 10 | 期: | 11 | 起(迄)頁: | 1839 | 來源出版物: | Vaccines | 摘要: | Recombinant proteins are essential in the development of subunit vaccines. In the design of many recombinant proteins, polyhistidine residues are added to the N- or C-termini of target sequences to facilitate purification. However, whether the addition of tag residues influences the immunogenicity of proteins remains unknown. In this study, the tag-free SARS-CoV-2 RBD and His-tag SARS-CoV-2 RBD proteins were investigated to determine whether there were any differences in their receptor binding affinity and immunogenicity. The results showed that the tag-free RBD protein had a higher affinity for binding with hACE2 receptors than His-tag RBD proteins (EC50: 1.78 µM vs. 7.51 µM). On day 21 after primary immunization with the proteins, the serum ELISA titers of immunized mice were measured and found to be 1:1418 for those immunized with tag-free RBD and only 1:2.4 for His-tag RBD. Two weeks after the booster dose, tag-free-RBD-immunized mice demonstrated a significantly higher neutralizing titer of 1:369 compared with 1:7.9 for His-tag-RBD-immunized mice. Furthermore, neutralizing antibodies induced by tag-free RBD persisted for up to 5 months and demonstrated greater cross-neutralization of the SARS-CoV-2 Delta variant. Evidence from Western blotting showed that the serum of His-tag-RBD-immunized mice recognized irrelevant His-tag proteins. Collectively, we conclude that the addition of a polyhistidine tag on a recombinant protein, when used as a COVID-19 vaccine antigen, may significantly impair protein immunogenicity against SARS-CoV-2. Antibody responses induced were clearly more rapid and robust for the tag-free SARS-CoV-2 RBD than the His-tag SARS-CoV-2 RBD. These findings provide important information for the design of antigens used in the development of COVID-19 subunit vaccines. |
URI: | https://scholars.lib.ntu.edu.tw/handle/123456789/626684 https://www.scopus.com/inward/record.uri?eid=2-s2.0-85141807312&doi=10.3390%2fvaccines10111839&partnerID=40&md5=bb6e72918db99c195e4a842297901f4d |
ISSN: | 2076-393X | DOI: | 10.3390/vaccines10111839 |
顯示於: | 生化科技學系 |
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