PMID- 15668999
OWN - NLM
STAT- MEDLINE
DCOM- 20050719
LR  - 20061115
IS  - 1615-9853 (Print)
IS  - 1615-9853 (Linking)
VI  - 5
IP  - 3
DP  - 2005 Feb
TI  - Identification of grass pollen allergens by two-dimensional gel electrophoresis 
      and serological screening.
PG  - 729-36
AB  - Approximately 50% of allergic patients are sensitized against grass pollen 
      allergens. The characterization of specific immunoglobulin E (IgE) reactivity to 
      allergen components in pollen-allergic patients is fundamental for clinical 
      diagnosis and for immunotherapy. Complex allergen extracts are commonly used in 
      diagnostic tests as well as in immunotherapy preparations, but their composition 
      in single allergenic molecules is only partially known. Diagnostic tests which 
      utilize recombinant or immuno-purified allergens have been made available in 
      clinical practice. They allow to obtain specific profiles of IgE reactivity, but 
      the panel of available molecules is far from complete. Here, we used a proteomic 
      approach in order to detect grass allergens from a natural protein extract. A 
      five-grass pollen extract used for diagnosis and immunotherapy was resolved by 
      two dimensional gel electrophoresis (2-DE), and assayed with 9 sera from 
      pollen-allergic patients whose sensitization profile was dissected by using IgE 
      reactivity to recombinant allergens. 2-DE immunoreactivity patterns were matched 
      with IgE reactivity to identify protein spots as candidate allergens. Identity 
      was confirmed by mass spectrometry analysis. We identified 6 out of 8 expected 
      clinically relevant allergens in the natural grass extract. Moreover, we 
      identified different molecular isoforms of single allergens, thus obtaining a 
      more detailed profile of IgE reactivity. Some discrepancies in protein isoform 
      profile and sera immunoreactivity between recombinant and native allergen 5 from 
      Phleum pratense were observed and a new putative allergen was described. The 
      proteomic approach applied to the analysis of a natural allergen allows the 
      comprehensive evaluation of the sensitization profile of allergic patients and 
      the identification of new allergens.
FAU - Corti, Valeria
AU  - Corti V
AD  - Proteome Biochemistry, San Raffaele Scientific Institute, Milan, Italy.
FAU - Cattaneo, Angela
AU  - Cattaneo A
FAU - Bachi, Angela
AU  - Bachi A
FAU - Rossi, Renato E
AU  - Rossi RE
FAU - Monasterolo, Giorgio
AU  - Monasterolo G
FAU - Paolucci, Clara
AU  - Paolucci C
FAU - Burastero, Samuele E
AU  - Burastero SE
FAU - Alessio, Massimo
AU  - Alessio M
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Germany
TA  - Proteomics
JT  - Proteomics
JID - 101092707
RN  - 0 (Allergens)
RN  - 0 (Plant Extracts)
RN  - 0 (Plant Proteins)
RN  - 0 (Protein Isoforms)
RN  - 37341-29-0 (Immunoglobulin E)
SB  - IM
MH  - Allergens/*analysis/blood/immunology
MH  - Chromatography, Liquid
MH  - Electrophoresis, Gel, Two-Dimensional
MH  - Humans
MH  - Immunoglobulin E/immunology
MH  - Mass Spectrometry/methods
MH  - Plant Extracts/chemistry
MH  - Plant Proteins/analysis/blood/immunology
MH  - Poaceae/*immunology
MH  - Pollen/*immunology
MH  - Protein Isoforms/analysis/blood/immunology
MH  - Proteomics
EDAT- 2005/01/26 09:00
MHDA- 2005/07/20 09:00
CRDT- 2005/01/26 09:00
PHST- 2005/01/26 09:00 [pubmed]
PHST- 2005/07/20 09:00 [medline]
PHST- 2005/01/26 09:00 [entrez]
AID - 10.1002/pmic.200401038 [doi]
PST - ppublish
SO  - Proteomics. 2005 Feb;5(3):729-36. doi: 10.1002/pmic.200401038.