High-throughgput phage-display screening in array format Articles uri icon

authors

  • Díez, P.
  • Jara-Acevedo, R.
  • González-González, M.
  • CASADO VELA, JUAN
  • Dasilva, N.
  • Lécrevisse, Q.
  • Bartolomé, R.
  • Claros, J.C.
  • GONZALEZ GONZALEZ, ALBERTO
  • Orfao, A.
  • LOPEZ JIMENEZ, RAQUEL
  • Fuentes, M.

publication date

  • January 2015

start page

  • 34

end page

  • 41

volume

  • 79-80

International Standard Serial Number (ISSN)

  • 0141-0229

abstract

  • Emerging technologies for the design and generation of human antibodies require improved approaches enabling their screening, characterization and validation. Currently, strategies based on ELISA or western blot are used to that aim. However, the ever increasing number of novel antibodies generated would benefit from the development of new high-throughput (HT) platforms facilitating rapid antibody identification and characterization. Herein, we describe a protein chip bearing recombinant phage particles and based on a large phage antibody library. In this paper we have set forth a novel implementation which provides a powerful and simple methodology enabling the identification of single-chain variable fragments (scFv). As a proof-of-principle of this method, we tested it with recombinant antigen (human recombinant interleukin 8). Additionally, we developed a novel bioinformatics tool that serves to compare this novel strategy with traditional methods. The method described here, together with associated informatics tools, is robust, relatively fast and represents a step-forward in protocols including phage library screenings. © 2015 Elsevier Inc..

keywords

  • antibody chip high-throughput screening phage library phage-display array bioinformatics recombinant proteins throughput bioinformatics tools chip emerging technologies high throughput screening phage display phage display screening phage libraries single chain variable fragments antibodies immunoglobulin heavy chain immunoglobulin kappa chain recombinant interleukin 8 single chain fragment variable antibody dna il8 protein; human immobilized protein interleukin 8 peptide library recombinant protein single chain fragment variable antibody antibody library article controlled study dna sequence heavy chain high throughput screening human hydrophobicity phage display polymerase chain reaction protein binding protein expression biology cell surface display chemistry genetics high throughput screening immunology molecular cloning molecular genetics nucleotide sequence peptide library procedures protein microarray base sequence cell surface display techniques cloning; molecular computational biology dna high-throughput screening assays humans immobilized proteins interleukin-8 molecular sequence data peptide library protein array analysis recombinant proteins single-chain antibodies