Selection and characterization of human scFvs targeting the SARS-CoV-2 nucleocapsid protein isolated from antibody libraries of COVID-19 patients.

Autor: Lisi S; Bio@SNS Laboratory, Scuola Normale Superiore, 56126, Pisa, Italy., Malerba F; Fondazione EBRI (European Brain Research Institute) Rita Levi-Montalcini, 00161, Rome, Italy., Quaranta P; Retrovirus Centre, Department of Translational Research, University of Pisa, 56126, Pisa, Italy.; Virology Operative Unit, Pisa University Hospital, 56124, Pisa, Italy., Florio R; Fondazione EBRI (European Brain Research Institute) Rita Levi-Montalcini, 00161, Rome, Italy., Vitaloni O; Bio@SNS Laboratory, Scuola Normale Superiore, 56126, Pisa, Italy., Monaca E; Structural Biology and Biophysics Unit, Fondazione Ri.MED, 90133, Palermo, Italy., Bruni Ercole B; Fondazione EBRI (European Brain Research Institute) Rita Levi-Montalcini, 00161, Rome, Italy., Bitonti AR; Bio@SNS Laboratory, Scuola Normale Superiore, 56126, Pisa, Italy., Del Perugia O; Bio@SNS Laboratory, Scuola Normale Superiore, 56126, Pisa, Italy., Mignanelli M; Bio@SNS Laboratory, Scuola Normale Superiore, 56126, Pisa, Italy., Perrera P; Retrovirus Centre, Department of Translational Research, University of Pisa, 56126, Pisa, Italy., Sabbatella R; Structural Biology and Biophysics Unit, Fondazione Ri.MED, 90133, Palermo, Italy., Raimondi F; Bio@SNS Laboratory, Scuola Normale Superiore, 56126, Pisa, Italy., Piazza CR; Retrovirus Centre, Department of Translational Research, University of Pisa, 56126, Pisa, Italy.; Department of Medical Biotechnologies, University of Siena, 53100, Siena, Italy., Moles A; Genomnia Srl, 20091, Bresso, MI, Italy.; Institute of Biochemistry and Cell Biology, CNR, 80131, Napoli, Italy., Alfano C; Structural Biology and Biophysics Unit, Fondazione Ri.MED, 90133, Palermo, Italy., Pistello M; Retrovirus Centre, Department of Translational Research, University of Pisa, 56126, Pisa, Italy.; Virology Operative Unit, Pisa University Hospital, 56124, Pisa, Italy., Cattaneo A; Bio@SNS Laboratory, Scuola Normale Superiore, 56126, Pisa, Italy. antonino.cattaneo@sns.it.; Fondazione EBRI (European Brain Research Institute) Rita Levi-Montalcini, 00161, Rome, Italy. antonino.cattaneo@sns.it.
Jazyk: angličtina
Zdroj: Scientific reports [Sci Rep] 2024 Jul 09; Vol. 14 (1), pp. 15864. Date of Electronic Publication: 2024 Jul 09.
DOI: 10.1038/s41598-024-66558-0
Abstrakt: In 2019, the novel SARS-CoV-2 coronavirus emerged in China, causing the pneumonia named COVID-19. At the beginning, all research efforts were focused on the spike (S) glycoprotein. However, it became evident that the nucleocapsid (N) protein is pivotal in viral replication, genome packaging and evasion of the immune system, is highly immunogenic, which makes it another compelling target for antibody development alongside the spike protein. This study focused on the construction of single chain fragments variable (scFvs) libraries from SARS-CoV-2-infected patients to establish a valuable, immortalized and extensive antibodies source. We used the Intracellular Antibody Capture Technology to select a panel of scFvs against the SARS-CoV-2 N protein. The whole panel of scFv was expressed and characterized both as intrabodies and recombinant proteins. ScFvs were then divided into 2 subgroups: those that exhibited high binding activity to N protein when expressed in yeast or in mammalian cells as intrabodies, and those purified as recombinant proteins, displaying affinity for recombinant N protein in the nanomolar range. This panel of scFvs against the N protein represents a novel platform for research and potential diagnostic applications.
(© 2024. The Author(s).)
Databáze: MEDLINE
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