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Original Article

IgE Binding Reactivity of Peptide Fragments of Bla g 4, a Major German Cockroach Allergen

Kwang Hyun Shin1, Kyoung Yong Jeong1, Chein-Soo Hong2, Tai-Soon Yong1
The Korean Journal of Parasitology 2009;47(1):31-36.
Published online: March 12, 2009

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Corresponding author (tsyong212@yuhs.ac)
• Received: July 29, 2008   • Revised: January 30, 2009   • Accepted: February 6, 2009

Copyright © 2009 by The Korean Society for Parasitology

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IgE Binding Reactivity of Peptide Fragments of Bla g 4, a Major German Cockroach Allergen
Korean J Parasitol. 2009;47(1):31-36.   Published online March 12, 2009
DOI: https://doi.org/10.3347/kjp.2009.47.1.31
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IgE Binding Reactivity of Peptide Fragments of Bla g 4, a Major German Cockroach Allergen
Korean J Parasitol. 2009;47(1):31-36.   Published online March 12, 2009
DOI: https://doi.org/10.3347/kjp.2009.47.1.31
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IgE Binding Reactivity of Peptide Fragments of Bla g 4, a Major German Cockroach Allergen
Image Image Image Image
Fig. 1 (A) Schematic representation of Bla g 4 fragments for epitope analysis. (B) SDS-PAGE of full-length and peptide fragments. Proteins run on 5-20% gradient SDS-PAGE gel and stained with Coomassie brilliant blue. M, Molecular weight markers in kDa; F, full-length; E1, 1-100 amino acid; E2, 34-77; E3, 74-117; E4, 114-156; and E5, 153-182.
Fig. 2 IgE reactivity of human sera against recombinant Bla g 4. The horizontal line indicates the cut-off value. ▴, sera from healthy controls; ♦, sera from German cockroach-sensitized subjects (Uni-CAP > 0.7 kU/L).
Fig. 3 (A) Binding profiles of IgE antibodies to recombinant Bla g 4 and relevant recombinant proteins by ELISA. 1-4, allergic sera; 5, pooled sera from healthy controls; 6, bovine serum albumin. The horizontal line indicates the cut-off value. (B) IgE-binding reactivities of peptide fragments of German cockroach allergen, Bla g 4.
Fig. 4 Sequence alignment of allergenic lipocalins: Bla g 4 (Blattella germanica, EF202172), Bos d 5 (Bos domesticus, X14712), Bos d 2 (B. domesticus, L42867), Equ c 1 (Equus caballus, U70823), Can f 1 (Canis familiaris, AF027177), Can f 2 (C. familiaris, AF027178), Mus m 1 (Mus musculus, X03208), Rat n 1 (Rattus norvegicus, M26835). *, fully conserved residues; : , strongly conserved residues; ·, weakly conserved residues. The shaded areas indicate the IgE-binding regions.

Fig. 1

Fig. 2

Fig. 3

Fig. 4

IgE Binding Reactivity of Peptide Fragments of Bla g 4, a Major German Cockroach Allergen
Serum sample Sexa Age (year) Diagnosis OD450b P1 M 43 Allergic asthma 0.0795 P2 F 52 Dermatofibroma 0.0855 P3 F 45 Allergic rhinitis 0.0730 P4 F 57 Allergic asthma 0.0725 P5 F 46 Allergic rhinitis 0.0745 P6 M 49 Allergic asthma 0.0770 P7 F 22 Atopic dermatitis 0.0785 P8 M 35 Allergic rhinitis 0.0740 P9 M 49 Allergic asthma 0.0890c P10 M 48 Allergic asthma 0.0865 P11 M 11 Atopic dermatitis 0.2155 P12 F 45 Allergic asthma 0.0900 P13 M 26 Atopic dermatitis 0.0790 P14 M 45 Allergic rhinitis 0.0910 P15 M 56 Allergic asthma 0.0860 P16 M 47 Atopic dermatitis 0.1635 P17 F 37 Allergic asthma 0.0800 P18 F 58 Allergic asthma 0.0795 P19 M 52 Allergic asthma 0.0725 P20 M 38 Allergic rhinitis 0.0765 P21 M 26 Allergic asthma 0.0725 P22 M 33 Allergic rhinitis 0.0835 P23 F 17 Atopic dermatitis 0.0825 P24 F 36 Allergic asthma 0.0730 P25 M 30 Allergic rhinitis 0.0730 P26 M 7 Atopic dermatitis 0.0810 P27 M 20 Allergic asthma 0.0725 P28 M 24 Allergic asthma 0.1030 P29 M 41 Allergic asthma 0.0680 P30 M 14 Atopic dermatitis 0.0690 P31 F 15 Atopic dermatitis 0.1045 P32 M 53 Allergic rhinitis 0.0680 Oligonucleotides Sequencesa Bg4 -E1F 5′-GGATCCGGCAGTTTTGGCACTA-3′ Bg4 -E1R 5′-CTCGAGGTGCTGTATCATTTCT-3′ Bg4 -E2F 5′-GGATCCGTGGATTATTGCAGCC-3′ Bg4 -E2R 5′-CTCGAGTCTTATAGCAGTCTTA-3′ Bg4 -E3F 5′-GGATCCGGCTATAAGAGGACGA-3′ Bg4 -E3R 5′-CTCGAGGCCTTCCACAATTGCG-3′ Bg4 -E4F 5′-GGATCCAGTGGAAGGCTGTCCC-3′ Bg4 -E4R 5′-CTCGAGCACCTGATCCAAAGTG-3′ Bg4 -E5F 5′-GGATCCGGATCAGGTGAATCAA-3′ Bg4 -E5R 5′-CTCGAGTTAGTTACATGTGGAG-3′
Table 1. IgE reactivity of human sera against recombinant Bla g 4

M, male; F, female;

The cut-off value is 0.086419;

Positives are indicated by underlines.

Table 2. Sequences of oligonucleotides used for the production of fragments of Bla g 4

Underline indicates restriction enzyme sites.

Table 1.

Table 2.

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