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Tapping allergen repertoires by advanced cloning technologies.

R Crameri1, R Kodzius, Z Konthur

  • 1Swiss Institute of Allergy and Asthma Research (SIAF), Davos, Switzerland. crameri@siaf.unizh.ch

International Archives of Allergy and Immunology
|April 18, 2001
PubMed
Summary

This study introduces a rapid method for identifying and characterizing allergens from various sources. By combining IgE-based selection with robotic screening, researchers can efficiently clone and analyze the complete allergenic repertoire, advancing allergy diagnostics and therapeutics.

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Area of Science:

  • Molecular biology
  • Immunology
  • Biotechnology

Background:

  • Complex allergenic sources contain numerous IgE-binding molecules.
  • Characterizing these molecules is crucial for developing accurate allergy diagnostics.
  • Recombinant allergens are needed to mimic whole allergen extracts.

Purpose of the Study:

  • To develop a fast and cost-effective method for isolating and characterizing a wide range of allergens.
  • To enable the cloning and analysis of the entire allergenic repertoire from diverse sources.

Main Methods:

  • Utilizing phage surface display of cDNA libraries enriched for allergen-expressing clones.
  • Employing IgE from allergic patients for selective enrichment (biopanning).
  • Implementing automated robotic technology for high-throughput screening and clone identification.

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Main Results:

  • Successfully identified numerous different allergen sequences from various sources including fungi, food, and human tissues.
  • Demonstrated rapid cloning and characterization of the whole allergenic repertoire within weeks.
  • Confirmed high probability of identified cDNAs encoding allergens.

Conclusions:

  • Robotic-based high-throughput screening is effective for characterizing IgE-binding clones.
  • This approach allows rapid identification of diverse allergens from complex sources.
  • The method facilitates comprehensive allergen repertoire analysis for improved allergy research.