A recombinant functional variant of the olive pollen allergen Ole e 10 expressed in baculovirus system.
Pollens have been reported as important sources of antigens causing type-I allergy and, among them, olive pollen has high clinical relevance in Mediterranean countries. The most recently described olive allergen, Ole e 10, is involved in cross-reactivity phenomena and related to asthma induction in allergic patients. These immunologic features make this allergen a good candidate to be included in diagnosis and therapy of protocols of allergic diseases. Since the availability of Ole e 10 from the olive pollen is limited, the allergen has been efficiently expressed in the baculovirus/insect cell system. The Ole e 10-cDNA inserted into the transfer vector pBacPAK8 allowed the expression of the recombinant protein in cultured Sf21 cells. Recombinant Ole e 10 (rOle e 10) was purified from the culture after dialysis and three chromatographic steps. Mass spectrometry, Edman degradation, IgE- and IgG-binding analyses were employed to characterize the recombinant allergen, which showed molecular and immunological equivalence with the natural protein. Affinity gel electrophoresis in presence of laminarin (1,3-beta-glucan) revealed that rOle e 10 retains identical carbohydrate-binding capacity than the natural allergen. In conclusion, the recombinant expression of Ole e 10 in baculovirus/insect cell system produces a homogeneous and biologically active allergen that could be useful for clinical and scientific purposes.
Barral
Patricia
P
Departamento de Bioquímica y Biología Molecular I, Facultad de Químicas, Universidad Complutense, 28040 Madrid, Spain.
Serrano
Alicia G
AG
Batanero
Eva
E
Pérez-Gil
Jesús
J
Villalba
Mayte
M
Rodríguez
Rosalía
R
D000485:Allergens / Q000276:immunology*; D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D015153:Blotting, Western; D002478:Cells, Cultured; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D014644:Genetic Variation; D006801:Humans; D006967:Hypersensitivity / Q000209:etiology / Q000503:physiopathology; D007073:Immunoglobulin E / Q000276:immunology; D007313:Insecta / Q000166:cytology; D013058:Mass Spectrometry; D008970:Molecular Weight; D031658:Olea / Q000235:genetics / Q000276:immunology*; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D011058:Pollen / Q000276:immunology*; D011994:Recombinant Proteins / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification
D000485:Allergens; D010940:Plant Proteins; D011994:Recombinant Proteins; D007073:Immunoglobulin E