Extraction and quantification of Ole e 1 from atmospheric air samples: An optimized protocol

Area of Science:

• Allergy and Immunology
• Environmental Science
• Biochemistry

Background:

• Olive pollen (Olea europaea) is a primary cause of pollinosis in Mediterranean regions.
• Quantifying the major allergen Ole e 1 typically uses Enzyme-Linked ImmunoSorbent Assay (ELISA), but protein extraction methods are often underspecified.
• Existing ELISA protocols lack comparative analysis of different extraction buffers.

Purpose of the Study:

• To develop an optimized and reproducible ELISA protocol for quantifying atmospheric Ole e 1.
• To evaluate the impact of different buffer compositions and antibody concentrations on protein extraction and allergen quantification.
• To establish a reliable method for monitoring Ole e 1 levels in ambient air.

Main Methods:

• Atmospheric Ole e 1 allergen and olive pollen concentrations were monitored using an automatic multi-vial cyclonic sampler and a Hirst volumetric pollen trap, respectively.
• ELISA protocols were optimized by varying capture/detection antibody concentrations and evaluating different extraction buffers.
• Protein extraction efficiency was compared across various buffer modifications.

Main Results:

• A modified buffer significantly enhanced protein extraction efficiency for Ole e 1.
• Optimized ELISA protocol demonstrated highly significant correlations between daily pollen counts and allergen levels.
• The refined ELISA method reduced antibody requirements and enabled Ole e 1 detection even at very low or zero airborne pollen concentrations.

Conclusions:

• The developed ELISA protocol and optimized buffer significantly improve the quantification of atmospheric Ole e 1.
• This method offers a more sensitive and cost-effective approach for monitoring olive pollen allergen exposure.
• The findings contribute to better understanding and management of olive pollen-induced allergies.