A Proteomic Style Approach To Characterize a Grass Mix Product Reveals Potential Immunotherapeutic Benefit

World Allergy Organization Journal

Murray Skinner¹  Wemimo Alawode¹  Nicola Swan¹  Alan Bullimore¹ 

[1] Allergy Therapeutics, Dominion Way, Worthing, West Sussex, 8SA, UK

关键词: mass spectrometry;    homology and standardization;    extraction;    characterization;    allergens;   

Others  :  1145541
DOI  :  10.1097/WOX.0b013e3182260bd2

【 摘 要 】

Background

Grass allergy immunotherapies often consist of a mix of different grass extracts, each containing several proteins of different physiochemical properties; however, the subtle contributions of each protein are difficult to elucidate. This study aimed to identify and characterize the group 1 and 5 allergens in a 13 grass extract and to standardize the extraction method.

Methods

The grass pollens were extracted in isolation and pooled and also in combination and analyzed using a variety of techniques including enzyme-linked immunosorbent assay, liquid chromatography-mass spectrometry, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

Results

Gold-staining and IgE immunoblotting revealed a high degree of homology of protein bands between the 13 species and the presence of a densely stained doublet at 25-35 kD along with protein bands at approximately 12.5, 17, and 50 kD. The doublet from each grass species demonstrated a high level of group 1 and 5 interspecies homology. However, there were a number of bands unique to specific grasses consistent with evolutionary change and indicative that a grass mix immunotherapeutic could be considered broad spectrum.

Conclusions

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and IgE immunoblotting showed all 13 grasses share a high degree of homology, particularly in terms of group 1 and 5 allergens. IgE and IgG enzyme-linked immunosorbent assay potencies were shown to be independent of extraction method.

【 授权许可】

   
2011 World Allergy Organization; licensee BioMed Central Ltd.

【 图 表 】

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