Characterization and improvement of apple extracts for the diagnosis of apple IgE-mediated allergy

Ann Allergy Asthma Immunol. 2005 Nov;95(5):462-7. doi: 10.1016/S1081-1206(10)61172-6.

Abstract

Background: The production process of reliable fruit extracts is not well established.

Objectives: To improve the overall quality of apple extracts by reducing protein loss during the manufacturing process and to evaluate the improved extracts using in vivo and in vitro experiments.

Methods: Two types of extracts were prepared from peels of Golden Delicious apples (Malus domesticus). Extract A was extracted, 1:2 wt/vol, for 30 minutes at 40 degrees C in 0.01 M phosphate-buffered saline, and extract B was extracted, 1:2 wt/vol, in phosphate-buffered saline with 20% polyvinylpolypyrrolidone and 2-mmol/L EDTA. Both extracts were filtered, dialyzed in 3.5-kDa dialysis membranes, and lyophilized. The antigenic and allergenic profiles were analyzed using immunoblot and enzyme-linked immunosorbent assay. Nine patients clinically sensitive to apples and 12 controls underwent skin testing with both extracts.

Results: Extracts A and B had dry weight yields of 0.71% and 1.86% and protein contents of 104.6 and 257 microg/mg of freeze-dried material, respectively. A steady and progressive loss of protein, greater in extract A than in extract B, was detected at different intervals during the manufacturing process of both extracts. Extract B produced larger wheal sizes than extract A (P = .008). Enzyme-linked immunosorbent assay inhibition results confirmed that extract B had a greater inhibition capacity than extract A.

Conclusions: A progressive loss of protein content occurs during the manufacturing of apple extracts. Wheal sizes induced by extract B were significantly larger than those induced by extract A and prick-by-prick solutions. Extract B was also more potent in vitro than extract A.

Publication types

  • Comparative Study

MeSH terms

  • Adult
  • Allergens / chemistry*
  • Allergens / immunology
  • Female
  • Food Hypersensitivity / diagnosis*
  • Food Hypersensitivity / immunology
  • Fruit / chemistry*
  • Humans
  • Immunoglobulin E / blood
  • Immunoglobulin E / chemistry*
  • Male
  • Malus / chemistry*
  • Malus / immunology
  • Plant Extracts* / chemistry
  • Plant Extracts* / immunology
  • Proteins / analysis
  • Skin Tests
  • Spain

Substances

  • Allergens
  • Plant Extracts
  • Proteins
  • Immunoglobulin E