Purification and characterisation of a panel of peanut allergens suitable for use in allergy diagnosis.
Peanut is a major cause of type 1 hypersensitive reactions including anaphylaxis. This results from the presence of a number of protein allergens, six of which are being studied as part of the EU FP6 EuroPrevall programme. These are Ara h 1 (7S globulin), Ara h 2, Ara h 6 (2S albumins), Ara h 3/4 (11S globulins) and Ara h 8 (Bet v 1 homologue). Methods for the purification of Ara h 1, Ara h 3/4, Ara h 2 and Ara h 6 from peanut seeds and for the production of recombinant Ara h 8 in Escherichia coli are described with spectroscopic analyses being used to confirm that they are authentically folded. N-terminal sequencing of the proteins purified from peanut seeds also revealed details of the differences between isoforms and their generation by proteolytic processing within the seed. Preliminary IgE binding studies of the purified allergens confirmed that they retained their immunological properties indicating their suitability for use in allergy diagnosis.
Marsh
Justin
J
Rothamsted Research, Harpenden, Hertfordshire, UK.
Rigby
Neil
N
Wellner
Klaus
K
Reese
Gerald
G
Knulst
André
A
Akkerdaas
Jaap
J
van Ree
Ronald
R
Radauer
Christian
C
Lovegrove
Alison
A
Sancho
Ana
A
Mills
Clare
C
Vieths
Stefan
S
Hoffmann-Sommergruber
Karin
K
Shewry
Peter R
PR
D000485:Allergens / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*; D000595:Amino Acid Sequence; D010367:Arachis / Q000276:immunology*; D001483:Base Sequence; D006801:Humans; D007073:Immunoglobulin E / Q000276:immunology; D008969:Molecular Sequence Data; D021183:Peanut Hypersensitivity / Q000175:diagnosis*; D017433:Protein Structure, Secondary
D000485:Allergens; D007073:Immunoglobulin E