PMID- 14730563
OWN - NLM
STAT- MEDLINE
DCOM- 20040909
LR  - 20040119
IS  - 0173-0835 (Print)
IS  - 0173-0835 (Linking)
VI  - 25
IP  - 1
DP  - 2004 Jan
TI  - Analytical and preparative native polyacrylamide gel electrophoresis: 
      Investigation of the recombinant and natural major grass pollen allergen Phl p 2.
PG  - 14-9
AB  - Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western 
      blot are amongst the most popular methods for allergen characterization, such as 
      comparison of recombinant allergens with their natural counterparts. Native PAGE 
      was evaluated as a possible robust and simple method offering high-resolution 
      capacity for characterization of the major grass pollen allergen Phl p 2. 
      Analytical separation of recombinant Phl p 2 provided a superior quality control 
      in terms of homogeneity and, after Western blotting, immunoglobulin E (IgE) 
      reactivity. Separation of natural Phl p 2 identified two major isoforms which 
      were shown to have different N-terminal sequences and IgE-binding properties. 
      After isolation using preparative native PAGE in combination with 
      electrodialysis, both isoforms were investigated by specific proteolysis and 
      reversed-phase high-performance liquid chromatography (RP-HPLC). The results 
      demonstrate differences in the primary structures and that the recombinant 
      counterpart corresponds exactly to one isoform. Analytical and preparative native 
      PAGE thus proved to be powerful tools for the investigation of allergen isoforms 
      and quality control of recombinant counterparts.
FAU - Suck, Roland
AU  - Suck R
AD  - Allergopharma Joachim Ganzer KG, Reinbek, Germany. roland.suck@allergopharma.de
FAU - Petersen, Arnd
AU  - Petersen A
FAU - Weber, Bernhard
AU  - Weber B
FAU - Becker, Wolf-Meinhard
AU  - Becker WM
FAU - Fiebig, Helmut
AU  - Fiebig H
FAU - Cromwell, Oliver
AU  - Cromwell O
LA  - eng
PT  - Journal Article
PL  - Germany
TA  - Electrophoresis
JT  - Electrophoresis
JID - 8204476
RN  - 0 (Allergens)
RN  - 0 (PHLPII protein, Phleum pratense)
RN  - 0 (Plant Proteins)
RN  - 0 (Protein Isoforms)
RN  - 0 (Recombinant Proteins)
RN  - 37341-29-0 (Immunoglobulin E)
SB  - IM
MH  - Allergens/analysis/chemistry/*isolation & purification
MH  - Amino Acid Sequence
MH  - Antibody Affinity
MH  - Chromatography, High Pressure Liquid
MH  - Electrophoresis, Polyacrylamide Gel/*methods
MH  - Escherichia coli/genetics
MH  - Immunoglobulin E
MH  - Molecular Sequence Data
MH  - Plant Proteins/analysis/chemistry/*isolation & purification
MH  - Pollen/*immunology
MH  - Protein Isoforms/chemistry/isolation & purification
MH  - Recombinant Proteins
MH  - Sequence Alignment
EDAT- 2004/01/20 05:00
MHDA- 2004/09/10 05:00
CRDT- 2004/01/20 05:00
PHST- 2004/01/20 05:00 [pubmed]
PHST- 2004/09/10 05:00 [medline]
PHST- 2004/01/20 05:00 [entrez]
AID - 10.1002/elps.200305697 [doi]
PST - ppublish
SO  - Electrophoresis. 2004 Jan;25(1):14-9. doi: 10.1002/elps.200305697.