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Biomed Res Int. 2015;2015:703213. doi: 10.1155/2015/703213. Epub 2015 Nov 15.

One-Step Recovery of scFv Clones from High-Throughput Sequencing-Based Screening of Phage Display Libraries Challenged to Cells Expressing Native Claudin-1.

Author information

1
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.
2
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.
3
CEINGE Biotecnologie Avanzate S.C. a R.L., Via G. Salvatore 486, 80145 Napoli, Italy.

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.

PMID:
26649313
PMCID:
PMC4662980
DOI:
10.1155/2015/703213
[Indexed for MEDLINE]
Free PMC Article

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