The purification and characterisation of allergenic hazelnut seed proteins.
A lipid transfer protein (LTP, Cor a 8) together with the 11S (Cor a 9) and 7S seed storage globulins (Cor a 11) are major food allergens present in hazelnut. Methods are described for their purification and characterisation using in-gel tryptic digestion mass spectrometry to confirm their identities and circular dichroism and Fourier-transform infrared spectroscopies to demonstrate that they are authentically folded. Preliminary immunochemical studies have also confirmed that the purified preparations retain their immunological properties in terms of immunoglobulin E binding, determined by immunoblotting using serum from hazelnut allergic patients. These preparations form a basis for development of improved methods of diagnosis of food allergy based on the concept of component-resolved diagnosis.
Rigby
Neil M
NM
Institute of Food Research, Norwich, UK.
Marsh
Justin
J
Sancho
Ana I
AI
Wellner
Klaus
K
Akkerdaas
Jaap
J
van Ree
Ronald
R
Knulst
Andre
A
Fernández-Rivas
Montserrat
M
Brettlova
Vlasta
V
Schilte
Piet P
PP
Summer
Colin
C
Pumphrey
Richard
R
Shewry
Peter R
PR
Mills
E N Clare
EN
D000485:Allergens / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*; D000595:Amino Acid Sequence; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002942:Circular Dichroism; D031211:Corylus / Q000276:immunology*; D006801:Humans; D015151:Immunoblotting; D007073:Immunoglobulin E / Q000378:metabolism; D008969:Molecular Sequence Data; D021184:Nut Hypersensitivity / Q000209:etiology*; D010940:Plant Proteins / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*; D017433:Protein Structure, Secondary; D012639:Seeds / Q000737:chemistry / Q000276:immunology*; D021241:Spectrometry, Mass, Electrospray Ionization; D017550:Spectroscopy, Fourier Transform Infrared
D000485:Allergens; D010940:Plant Proteins; D007073:Immunoglobulin E