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Antigens Involved in Adaptive Immunity01:26

Antigens Involved in Adaptive Immunity

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Humanized Mediator Release Assay as a Read-Out for Allergen Potency
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Characterization of allergoids.

Martin Himly1, Jerónimo Carnés, Enrique Fernández-Caldas

  • 1Abt. für Molekulare Biologie, Universität Salzburg.

Arbeiten Aus Dem Paul-Ehrlich-Institut (Bundesinstitut Fur Impfstoffe Und Biomedizinische Arzneimittel) Langen/Hessen
|August 31, 2010
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Summary
This summary is machine-generated.

Physicochemical methods now allow for the characterization of allergoids, determining their molecular size and allergen composition. This advancement supports the development of alternative testing methods, reducing animal use in quality control.

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Area of Science:

  • Allergen immunotherapy
  • Analytical chemistry
  • Biochemistry

Background:

  • Characterizing allergoids, chemically modified allergens, has been challenging.
  • Previous methods struggled to quantify major allergens and detect minor allergens in allergoids.
  • Animal immunization was often required to generate antibodies for allergen detection.

Purpose of the Study:

  • To adapt and employ physicochemical methods for allergoid characterization.
  • To determine the molecular size and allergen composition of allergoids.
  • To provide alternative methods to animal testing for allergoid quality control.

Main Methods:

  • Online-HPSEC light scattering
  • Dynamic Light Scattering (DLS)
  • Analysis of allergen composition in birch pollen and D. pteronyssinus allergoids

Main Results:

  • Two-thirds of depigmented birch pollen allergoids had a molecular weight (MW) between 1000-2000 kDa.
  • Reproducibility analysis showed 73-77% polymerized molecules in P. pratense allergoid batches.
  • Allergen composition analysis revealed the presence of key allergens in B. alba and D. pteronyssinus allergoids, aligning with native extracts.

Conclusions:

  • Physicochemical methods effectively elucidate allergoid molecular size and composition.
  • The findings support the use of these methods as alternatives to animal testing in allergoid quality control.
  • This approach aligns with the European Directorate for Quality of Medicines and Healthcare's (EDQM) 3R principles for reducing animal use.