Recombinant allergens Pru av 1 and Pru av 4 and a newly identified lipid transfer protein in the in vitro diagnosis of cherry allergy Stephan Scheurer,

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Presentation transcript:

Recombinant allergens Pru av 1 and Pru av 4 and a newly identified lipid transfer protein in the in vitro diagnosis of cherry allergy Stephan Scheurer, PhDa, Elide A. Pastorello, MDb, Andrea Wangorsch, BSca, Marion Kästner, a, Dieter Haustein, PhDa, Stefan Vieths, PhDa Journal of Allergy and Clinical Immunology Volume 107, Issue 4, Pages 724-731 (April 2001) DOI: 10.1067/mai.2001.113864 Copyright © 2001 Mosby, Inc. Terms and Conditions

Fig. 1 Amino acid sequence comparison of LTP from cherry (Pru av 3), with allergenic LTPs denominated according the IUIS allergen nomenclature. The identified signal peptide and cysteines in disulfide bounds are underlined. Dots indicate identical amino acids. Acc.Nr. , Accession number. Journal of Allergy and Clinical Immunology 2001 107, 724-731DOI: (10.1067/mai.2001.113864) Copyright © 2001 Mosby, Inc. Terms and Conditions

Figure 2 Expression and purification of recombinant LTP (rLTP) cherry fusion protein in BL21(DE3) RP codon plus cells. Lane 1 , Molecular weight marker; lane 2 , BL21(DE3) RP without isopropyl-D-thiogalactopyranoside, overnight, 37°C; lane 3 , BL21(DE3) RP codon plus cells with 1 mmol/L isopropyl-D-thiogalactopyranoside, overnight, 37°C; lane 4 , recombinant LTP cherry fusion protein purified with nickel-nitrilotriacetic acid/agarose. Journal of Allergy and Clinical Immunology 2001 107, 724-731DOI: (10.1067/mai.2001.113864) Copyright © 2001 Mosby, Inc. Terms and Conditions

Fig. 3 Immunoblot inhibition of patient IgE (sera 10, 29, and 32; 1:10 dilution) to cherry peel extract (20 μg/cm slot) on the solid phase by means of preincubation with 15 μg pf cherry recombinant LTP (fusion-protein) (lane 2) , 100 μg of birch pollen extract (lane 3) , and 100 μg of cherry peel extract (lane 4) and without inhibitor (lane 1) . Serum from a nonallergic subject (C) was used as control. Immunodetection was performed with a mouse-anti human IgE alkaline phosphatase–labeled mAb (1:1000). CCD, Cross-reactive carbohydrate determinants. Journal of Allergy and Clinical Immunology 2001 107, 724-731DOI: (10.1067/mai.2001.113864) Copyright © 2001 Mosby, Inc. Terms and Conditions