tetano
Editor, Senior Moderator
J Med Virol
. 2024 Jan;96(1):e29425.
doi: 10.1002/jmv.29425. Generation of SARS-CoV-2 spike receptor binding domain mutants and functional screening for immune evaders using a novel lentivirus-based system
Junli Jia[SUP] 1 [/SUP], Emanuela Garbarino[SUP] 1 [/SUP], Yuhang Wang[SUP] 1 2 [/SUP], Jiaming Li[SUP] 1 [/SUP], Minmin Song[SUP] 1 [/SUP], Xin Zhang[SUP] 1 [/SUP], Xinjie Wang[SUP] 1 [/SUP], Lingyun Li[SUP] 3 [/SUP], Jing Chi[SUP] 4 [/SUP], Lunbiao Cui[SUP] 5 [/SUP], Huamin Tang[SUP] 1 6 [/SUP]
Affiliations
The emergence of rapid and continuous mutations of severe acute respiratory syndrome 2 (SARS-CoV-2) spike glycoprotein that increased with the Omicron variant points out the necessity to anticipate such mutations for conceiving specific and adaptable therapies to avoid another pandemic. The crucial target for the antibody treatment and vaccine design is the receptor binding domain (RBD) of the SARS-CoV-2 spike. It is also the site where the virus has shown its high ability to mutate and consequently escape immune response. We developed a robust and simple method for generating a large number of functional SARS-CoV-2 spike RBD mutants by error-prone PCR and a novel nonreplicative lentivirus-based system. We prepared anti-RBD wild type (WT) polyclonal antibodies and used them to screen and select for mutant libraries that escape inhibition of virion entry into recipient cells expressing human angiotensin-converting enzyme 2 and transmembrane serine protease 2. We isolated, cloned, and sequenced six mutants totally bearing nine mutation sites. Eight mutations were found in successive WT variants, including Omicron and other recombinants, whereas one is novel. These results, together with the detailed functional analyses of two mutants provided the proof of concept for our approach.
Keywords: RBD; SARS-CoV-2; immune-evasion; lentivirus.
. 2024 Jan;96(1):e29425.
doi: 10.1002/jmv.29425. Generation of SARS-CoV-2 spike receptor binding domain mutants and functional screening for immune evaders using a novel lentivirus-based system
Junli Jia[SUP] 1 [/SUP], Emanuela Garbarino[SUP] 1 [/SUP], Yuhang Wang[SUP] 1 2 [/SUP], Jiaming Li[SUP] 1 [/SUP], Minmin Song[SUP] 1 [/SUP], Xin Zhang[SUP] 1 [/SUP], Xinjie Wang[SUP] 1 [/SUP], Lingyun Li[SUP] 3 [/SUP], Jing Chi[SUP] 4 [/SUP], Lunbiao Cui[SUP] 5 [/SUP], Huamin Tang[SUP] 1 6 [/SUP]
Affiliations
- PMID: 38258313
- DOI: 10.1002/jmv.29425
The emergence of rapid and continuous mutations of severe acute respiratory syndrome 2 (SARS-CoV-2) spike glycoprotein that increased with the Omicron variant points out the necessity to anticipate such mutations for conceiving specific and adaptable therapies to avoid another pandemic. The crucial target for the antibody treatment and vaccine design is the receptor binding domain (RBD) of the SARS-CoV-2 spike. It is also the site where the virus has shown its high ability to mutate and consequently escape immune response. We developed a robust and simple method for generating a large number of functional SARS-CoV-2 spike RBD mutants by error-prone PCR and a novel nonreplicative lentivirus-based system. We prepared anti-RBD wild type (WT) polyclonal antibodies and used them to screen and select for mutant libraries that escape inhibition of virion entry into recipient cells expressing human angiotensin-converting enzyme 2 and transmembrane serine protease 2. We isolated, cloned, and sequenced six mutants totally bearing nine mutation sites. Eight mutations were found in successive WT variants, including Omicron and other recombinants, whereas one is novel. These results, together with the detailed functional analyses of two mutants provided the proof of concept for our approach.
Keywords: RBD; SARS-CoV-2; immune-evasion; lentivirus.