title: Purification and recombinant expression of major peanut allergen Ara h 1.
abstract: Reaction to peanut, as one of the major food allergens, has become an increasingly common life-threatening disorder. Although peanut allergens have been extensively identified, Ara h 1 is still too expensive to be applied in food safety or clinical utility. In this study, the purification, expression, and immunological analyses of Ara h 1 are investigated. It was shown that a high purity (>95%) of Ara h 1 could be prepared by either purification or expression. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), Western blot, and mass spectroscopy were used to identify the Ara h 1, and it was found that natural Ara h 1 (nAra h 1) and expressed Ara h 1 (rAra h 1) have the same properties, including amino acid sequence. In particular, rAra h 1 reacted positively with anti-nAra h 1 serum, showing their similar immunological property. Thus, by either purification or expression, Ara h 1 could be prepared with low cost, as performed in the present work. SDS-PAGE, mag trix-assisted laser desorption/ionization time-of-flight mass spectroscopy (MALDI-TOF MS), and immunological analysis confirmed that both forms of Ara h 1 had the same properties.
authors.affiliation: a State Key Laboratory of Food Science and Technology , Nanchang University , Nanchang , P.R. China.
authors.forename: Zhihua
authors.initials: Z
authors.lastname: Wu
authors.forename: Fei
authors.initials: F
authors.lastname: Yan
authors.forename: Xiaonan
authors.initials: X
authors.lastname: Wei
authors.forename: Xin
authors.initials: X
authors.lastname: Li
authors.forename: Ping
authors.initials: P
authors.lastname: Tong
authors.forename: Anshu
authors.initials: A
authors.lastname: Yang
authors.forename: Ronghua
authors.initials: R
authors.lastname: Tang
authors.forename: Hongbing
authors.initials: H
authors.lastname: Chen
mesh_terms: D000818:Animals; D052179:Antigens, Plant / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification*; D015153:Blotting, Western; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006023:Glycoproteins / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification*; D008297:Male; D008565:Membrane Proteins; D010940:Plant Proteins / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification*; D011817:Rabbits; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D052179:Antigens, Plant; C500889:Ara h 1 protein, Arachis hypogaea; D006023:Glycoproteins; D008565:Membrane Proteins; D010940:Plant Proteins; D011994:Recombinant Proteins
keywords: Ara h 1; MALDI-TOF MS; expression; immunological analysis; peanut; purification