Purification of allergenic proteins from peanut for preparative of the reactive solid phase of a specific IgE radioimmunoassay.
Peanut is one of the most allergenic foods. Detection of specific IgE in the serum of allergic patients requires the purification of allergenic proteins. In the present work, proteins were recovered from peanut kernel after successive treatment in acetone and diethyl ether. The proteins were dissolved in 0.05% TFA and analysed by RP-HPLC with a 0-100% gradient of methanol containing 0.05% TFA. The protein peaks were recovered and tested in SDS-PAGE. Eleven proteins were identified with a M(r) ranging from 13 to 81. Western blotting was performed with sera from allergic patients. Allergenic proteins had a M(r) of 15, 18, 19, 33, 41 and 67. By comparison, a protein fraction from peanut shell contained seven proteins with M(r) ranging from 15 to 81. Only two proteins with M(r) of 18 and 41 were detected in a Western blot. The protein fractions were coupled to epoxy-Sepharose and the gels were used as a solid reactive phase for detection by IgE-RIA of specific IgE from the serum of allergic patients.
Moutete
H F
HF
Laboratoire de Biochimie Cellulaire et Moléculaire en Nutrition, Faculté de Médecine, CHU de Nancy, France.
Olszewski
A
A
Gastin
I
I
Namour
F
F
Moneret-Vautrin
D A
DA
Guéant
J L
JL
D000485:Allergens / Q000302:isolation & purification*; D010367:Arachis; D015153:Blotting, Western; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D006967:Hypersensitivity / Q000097:blood / Q000276:immunology; D007073:Immunoglobulin E / Q000097:blood*; D010940:Plant Proteins / Q000302:isolation & purification*; D011863:Radioimmunoassay
D000485:Allergens; D010940:Plant Proteins; D007073:Immunoglobulin E