Construction of Human Naïve Antibody Gene Libraries

Michela Pasello, Alessandra Mallano, Michela Flego, Silvia Zamboni, Anna Maria Giudice, Katia Scotlandi

Research output: Contribution to journalArticlepeer-review

Abstract

Size and variability often represent an obstacle in generating an effective antibody gene library for the detection of an abundant repertoire of antigens. Therefore, optimizing the construction of a large library is essential for the selection of high-affinity reactive fragments. Here, we report a highly efficient method for the construction of a human naïve antibody gene library for the selection of antibodies as single-chain variable fragments. This protocol is based on many different sets of oligonucleotide primers and multistep electroporation and ligation reactions.This advanced method can be adopted by any molecular biology laboratory to generate a naïve library for use in isolating single-chain fragment variables against specific targets.

Original languageEnglish
Pages (from-to)73-91
Number of pages19
JournalMethods in molecular biology (Clifton, N.J.)
Volume1827
DOIs
Publication statusPublished - 2018

Keywords

  • Antibodies/genetics
  • Gene Library
  • Genetic Techniques
  • Humans
  • Leukocytes, Mononuclear/metabolism
  • Peptide Library
  • Single-Chain Antibodies/genetics

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