title: One-Step Recovery of scFv Clones from High-Throughput Sequencing-Based Screening of Phage Display Libraries Challenged to Cells Expressing Native Claudin-1.
abstract: Expanding the availability of monoclonal antibodies interfering with hepatitis C virus infection of hepatocytes is an active field of investigation within medical biotechnologies, to prevent graft reinfection in patients subjected to liver transplantation and to overcome resistances elicited by novel antiviral drugs. In this paper, we describe a complete pipeline for screening of phage display libraries of human scFvs against native Claudin-1, a tight-junction protein involved in hepatitis C virus infection, expressed on the cell surface of human hepatocytes. To this aim, we implemented a high-throughput sequencing approach for library screening, followed by a simple and effective strategy to recover active binder clones from enriched sublibraries. The recovered clones were successfully converted to active immunoglobulins, thus demonstrating the effectiveness of the whole procedure. This novel approach can guarantee rapid and cheap isolation of antibodies for virtually any native antigen involved in human diseases, for therapeutic and/or diagnostic applications.
authors.affiliation: Dipartimento di Medicina Molecolare e Biotecnologie Mediche, Università degli Studi di Napoli Federico II, Via S. Pansini 5, 80131 Napoli, Italy ; CEINGE Biotecnologie Avanzate S.C. a R.L., Via G. Salvatore 486, 80145 Napoli, Italy ; Associazione Culturale DiSciMuS RFC, 80026 Casoria, Italy.
authors.forename: Emanuele
authors.initials: E
authors.lastname: Sasso
authors.affiliation: Dipartimento di Medicina Molecolare e Biotecnologie Mediche, Università degli Studi di Napoli Federico II, Via S. Pansini 5, 80131 Napoli, Italy ; CEINGE Biotecnologie Avanzate S.C. a R.L., Via G. Salvatore 486, 80145 Napoli, Italy.
authors.forename: Rolando
authors.initials: R
authors.lastname: Paciello
authors.affiliation: Dipartimento di Medicina Molecolare e Biotecnologie Mediche, Università degli Studi di Napoli Federico II, Via S. Pansini 5, 80131 Napoli, Italy ; CEINGE Biotecnologie Avanzate S.C. a R.L., Via G. Salvatore 486, 80145 Napoli, Italy.
authors.forename: Francesco
authors.initials: F
authors.lastname: D'Auria
authors.affiliation: Dipartimento di Medicina Molecolare e Biotecnologie Mediche, Università degli Studi di Napoli Federico II, Via S. Pansini 5, 80131 Napoli, Italy ; CEINGE Biotecnologie Avanzate S.C. a R.L., Via G. Salvatore 486, 80145 Napoli, Italy.
authors.forename: Gennaro
authors.initials: G
authors.lastname: Riccio
authors.affiliation: Dipartimento di Medicina Molecolare e Biotecnologie Mediche, Università degli Studi di Napoli Federico II, Via S. Pansini 5, 80131 Napoli, Italy ; CEINGE Biotecnologie Avanzate S.C. a R.L., Via G. Salvatore 486, 80145 Napoli, Italy.
authors.forename: Guendalina
authors.initials: G
authors.lastname: Froechlich
authors.affiliation: CEINGE Biotecnologie Avanzate S.C. a R.L., Via G. Salvatore 486, 80145 Napoli, Italy.
authors.forename: Riccardo
authors.initials: R
authors.lastname: Cortese
authors.affiliation: Dipartimento di Medicina Molecolare e Biotecnologie Mediche, Università degli Studi di Napoli Federico II, Via S. Pansini 5, 80131 Napoli, Italy ; CEINGE Biotecnologie Avanzate S.C. a R.L., Via G. Salvatore 486, 80145 Napoli, Italy.
authors.forename: Alfredo
authors.initials: A
authors.lastname: Nicosia
authors.affiliation: Dipartimento di Medicina Molecolare e Biotecnologie Mediche, Università degli Studi di Napoli Federico II, Via S. Pansini 5, 80131 Napoli, Italy ; CEINGE Biotecnologie Avanzate S.C. a R.L., Via G. Salvatore 486, 80145 Napoli, Italy.
authors.forename: Claudia
authors.initials: C
authors.lastname: De Lorenzo
authors.affiliation: Dipartimento di Medicina Molecolare e Biotecnologie Mediche, Università degli Studi di Napoli Federico II, Via S. Pansini 5, 80131 Napoli, Italy ; CEINGE Biotecnologie Avanzate S.C. a R.L., Via G. Salvatore 486, 80145 Napoli, Italy ; Associazione Culturale DiSciMuS RFC, 80026 Casoria, Italy.
authors.forename: Nicola
authors.initials: N
authors.lastname: Zambrano
mesh_terms: D062445:Claudin-1 / Q000737:chemistry / Q000378:metabolism*; D057809:HEK293 Cells; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D019151:Peptide Library; D057127:Single-Chain Antibodies / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
chemical_list: D062445:Claudin-1; D019151:Peptide Library; D057127:Single-Chain Antibodies