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Interactome-Seq: A Protocol for Domainome Library Construction, Validation and Selection by Phage Display and Next Generation Sequencing
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ORFeome Phage Display.

Jonas Zantow1, Gustavo Marçal Schmidt Garcia Moreira2, Stefan Dübel1

  • 1Institut für Biochemie, Biotechnologie und Bioinformatik, Abteilung Biotechnologie, Technische Universität Braunschweig, Spielmannstr. 7, 38106, Braunschweig, Germany.

Methods in Molecular Biology (Clifton, N.J.)
|November 9, 2017
PubMed
Summary
This summary is machine-generated.

ORFeome phage display screens proteomes to find immunogenic proteins. This technology identifies disease biomarkers and vaccine candidates, even from unculturable organisms.

Keywords:
AntigenBiomarker identificationGenomic librariesImmunogenic proteinsORFeome phage displayOligopeptide phage displayOpen reading frame (ORF) selectioncDNA libraries

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Area of Science:

  • Biotechnology
  • Immunology
  • Molecular Biology

Background:

  • Phage display is a powerful technique for protein-protein interaction studies.
  • Identifying immunogenic proteins is crucial for diagnostics and therapeutics.
  • Current methods may miss proteins expressed only during host-pathogen interactions or from unculturable microbes.

Purpose of the Study:

  • To introduce ORFeome phage display technology.
  • To provide protocols for identifying immunogenic proteins using phage display.
  • To highlight the utility of ORFeome phage display for discovering novel biomarkers and vaccine targets.

Main Methods:

  • Cloning randomly fragmented whole genomes or metagenomes into a phage-display vector.
  • Utilizing positive selection for open reading frames (ORFs) to enhance library quality.
  • Screening phage libraries with disease-related immunoglobulins from patient serum to identify immunogenic proteins.

Main Results:

  • ORFeome phage display enables efficient functional screening of entire proteomes and metaproteomes.
  • The technology successfully identifies immunogenic proteins, including those expressed only during host-pathogen interactions.
  • It can detect very low-expressed, small immunogens, and proteins from non-cultivable organisms.

Conclusions:

  • ORFeome phage display is a valuable tool for identifying immunogenic proteins.
  • Identified proteins serve as potential biomarkers for diagnostic assays and vaccine development.
  • This method overcomes limitations of traditional cultivation-based approaches.