title: Design and construction of a new human naïve single-chain fragment variable antibody library, IORISS1.
abstract: Human monoclonal antibodies are a powerful tool with increasingly successful exploitations and the single chain fragment variable format can be considered the building block for the implementation of more complex and effective antibody-based constructs. Phage display is one of the best and most efficient methods to isolate human antibodies selected from an efficient and variable phage display library. We report a method for the construction of a human naïve single-chain variable fragment library, termed IORISS1. Many different sets of oligonucleotide primers as well as optimized electroporation and ligation reactions were used to generate this library of 1.2×10(9) individual clones. The key difference is the diversity of variable gene templates, which was derived from only 15 non-immunized human donors. The method described here, was used to make a new human naïve single-chain fragment variable phage display library that represents a valuable source of diverse antibodies that can be used as research reagents or as a starting point for the development of therapeutics. Using biopanning, we determined the ability of IORISS1 to yield antibodies. The results we obtained suggest that, by using an optimized protocol, an efficient phage antibody library can be generated.
authors.affiliation: CRS Development of Biomolecular Therapies, Experimental Oncology Laboratory, Rizzoli Orthopaedic Institute, 40136 Bologna, Italy; PROMETEO Laboratory, STB, RIT Department, Rizzoli Orthopaedic Institute, 40136 Bologna, Italy. Electronic address: michela.pasello@ior.it.
authors.forename: Michela
authors.initials: M
authors.lastname: Pasello
authors.affiliation: PROMETEO Laboratory, STB, RIT Department, Rizzoli Orthopaedic Institute, 40136 Bologna, Italy; Section of Pharmacogenetics, Drug Resistance and Experimental Therapeutics, Department of Drug Research and Evaluation, Istituto Superiore di Sanità, 00161 Rome, Italy.
authors.forename: Silvia
authors.initials: S
authors.lastname: Zamboni
authors.affiliation: Section of Pharmacogenetics, Drug Resistance and Experimental Therapeutics, Department of Drug Research and Evaluation, Istituto Superiore di Sanità, 00161 Rome, Italy.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Mallano
authors.affiliation: Section of Pharmacogenetics, Drug Resistance and Experimental Therapeutics, Department of Drug Research and Evaluation, Istituto Superiore di Sanità, 00161 Rome, Italy.
authors.forename: Michela
authors.initials: M
authors.lastname: Flego
authors.affiliation: CRS Development of Biomolecular Therapies, Experimental Oncology Laboratory, Rizzoli Orthopaedic Institute, 40136 Bologna, Italy; PROMETEO Laboratory, STB, RIT Department, Rizzoli Orthopaedic Institute, 40136 Bologna, Italy.
authors.forename: Piero
authors.initials: P
authors.lastname: Picci
authors.affiliation: Section of Pharmacogenetics, Drug Resistance and Experimental Therapeutics, Department of Drug Research and Evaluation, Istituto Superiore di Sanità, 00161 Rome, Italy.
authors.forename: Maurizio
authors.initials: M
authors.lastname: Cianfriglia
authors.affiliation: CRS Development of Biomolecular Therapies, Experimental Oncology Laboratory, Rizzoli Orthopaedic Institute, 40136 Bologna, Italy; PROMETEO Laboratory, STB, RIT Department, Rizzoli Orthopaedic Institute, 40136 Bologna, Italy. Electronic address: katia.scotlandi@ior.it.
authors.forename: Katia
authors.initials: K
authors.lastname: Scotlandi
mesh_terms: D000911:Antibodies, Monoclonal / Q000235:genetics*; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D064368:Healthy Volunteers; D006801:Humans; D008214:Lymphocytes / Q000276:immunology; D009841:Oligonucleotides / Q000235:genetics; D019151:Peptide Library; D057127:Single-Chain Antibodies / Q000235:genetics*
chemical_list: D000911:Antibodies, Monoclonal; D017931:DNA Primers; D009841:Oligonucleotides; D019151:Peptide Library; D057127:Single-Chain Antibodies
keywords: Biopanning; IORISS1; Monoclonal antibody; Naïve library single-chain variable fragment; Phage display