Cargando…

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

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 characterise the group 1...

Descripción completa

Detalles Bibliográficos
Autores principales: Skinner, Murray, Bullimore, Alan, Swanc, Nicola, Alawode, Wemimo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: World Allergy Organization Journal 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3512597/
http://dx.doi.org/10.1097/01.WOX.0000411754.75035.ed
_version_ 1782251757151191040
author Skinner, Murray
Bullimore, Alan
Swanc, Nicola
Alawode, Wemimo
author_facet Skinner, Murray
Bullimore, Alan
Swanc, Nicola
Alawode, Wemimo
author_sort Skinner, Murray
collection PubMed
description 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 characterise the group 1 and 5 allergens in a 13 grass extract and to standardise the extraction method. METHODS: The grass pollens were extracted in isolation and pooled and also in combination and analysed using a variety of techniques including enzyme-linked immunosorbent assay (ELISA), liquid chromatography-mass spectrometry (LC-MS) and sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). 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 to 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: SDS-PAGE 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 ELISA potencies were shown to be independent of extraction method.
format Online
Article
Text
id pubmed-3512597
institution National Center for Biotechnology Information
language English
publishDate 2012
publisher World Allergy Organization Journal
record_format MEDLINE/PubMed
spelling pubmed-35125972012-12-21 9 A Proteomic Style Approach to Characterize a Grass Mix Product Reveals Potential Immunotherapeutic Benefit Skinner, Murray Bullimore, Alan Swanc, Nicola Alawode, Wemimo World Allergy Organ J Abstracts of the XXII World Allergy Congress 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 characterise the group 1 and 5 allergens in a 13 grass extract and to standardise the extraction method. METHODS: The grass pollens were extracted in isolation and pooled and also in combination and analysed using a variety of techniques including enzyme-linked immunosorbent assay (ELISA), liquid chromatography-mass spectrometry (LC-MS) and sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). 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 to 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: SDS-PAGE 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 ELISA potencies were shown to be independent of extraction method. World Allergy Organization Journal 2012-02-17 /pmc/articles/PMC3512597/ http://dx.doi.org/10.1097/01.WOX.0000411754.75035.ed Text en Copyright © 2012 by World Allergy Organization
spellingShingle Abstracts of the XXII World Allergy Congress
Skinner, Murray
Bullimore, Alan
Swanc, Nicola
Alawode, Wemimo
9 A Proteomic Style Approach to Characterize a Grass Mix Product Reveals Potential Immunotherapeutic Benefit
title 9 A Proteomic Style Approach to Characterize a Grass Mix Product Reveals Potential Immunotherapeutic Benefit
title_full 9 A Proteomic Style Approach to Characterize a Grass Mix Product Reveals Potential Immunotherapeutic Benefit
title_fullStr 9 A Proteomic Style Approach to Characterize a Grass Mix Product Reveals Potential Immunotherapeutic Benefit
title_full_unstemmed 9 A Proteomic Style Approach to Characterize a Grass Mix Product Reveals Potential Immunotherapeutic Benefit
title_short 9 A Proteomic Style Approach to Characterize a Grass Mix Product Reveals Potential Immunotherapeutic Benefit
title_sort 9 a proteomic style approach to characterize a grass mix product reveals potential immunotherapeutic benefit
topic Abstracts of the XXII World Allergy Congress
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3512597/
http://dx.doi.org/10.1097/01.WOX.0000411754.75035.ed
work_keys_str_mv AT skinnermurray 9aproteomicstyleapproachtocharacterizeagrassmixproductrevealspotentialimmunotherapeuticbenefit
AT bullimorealan 9aproteomicstyleapproachtocharacterizeagrassmixproductrevealspotentialimmunotherapeuticbenefit
AT swancnicola 9aproteomicstyleapproachtocharacterizeagrassmixproductrevealspotentialimmunotherapeuticbenefit
AT alawodewemimo 9aproteomicstyleapproachtocharacterizeagrassmixproductrevealspotentialimmunotherapeuticbenefit