Design and construction of a new human naïve single-chain fragment variable antibody library, IORISS1

Michela Pasello, Silvia Zamboni, Alessandra Mallano, Michela Flego, Piero Picci, Maurizio Cianfriglia, Katia Scotlandi

Research output: Contribution to journalArticle

5 Citations (Scopus)

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 × 109 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.

Original languageEnglish
Pages (from-to)1-11
Number of pages11
JournalJournal of Biotechnology
Volume224
DOIs
Publication statusPublished - Apr 20 2016

Fingerprint

Single-Chain Antibodies
Antibodies
Bacteriophages
Display devices
Libraries
DNA Primers
Monoclonal antibodies
Electroporation
Oligonucleotides
Ligation
Clone Cells
Genes
Monoclonal Antibodies
Research

Keywords

  • Biopanning
  • IORISS1
  • Monoclonal antibody
  • Naïve library single-chain variable fragment
  • Phage display

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology

Cite this

Design and construction of a new human naïve single-chain fragment variable antibody library, IORISS1. / Pasello, Michela; Zamboni, Silvia; Mallano, Alessandra; Flego, Michela; Picci, Piero; Cianfriglia, Maurizio; Scotlandi, Katia.

In: Journal of Biotechnology, Vol. 224, 20.04.2016, p. 1-11.

Research output: Contribution to journalArticle

Pasello, Michela ; Zamboni, Silvia ; Mallano, Alessandra ; Flego, Michela ; Picci, Piero ; Cianfriglia, Maurizio ; Scotlandi, Katia. / Design and construction of a new human naïve single-chain fragment variable antibody library, IORISS1. In: Journal of Biotechnology. 2016 ; Vol. 224. pp. 1-11.
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