Open Access

Enhanced sensitivity of capture IgE‑ELISA based on a recombinant Der f 1/2 fusion protein for the detection of IgE antibodies targeting house dust mite allergens

  • Authors:
    • Zhen Zhang
    • Zelang Cai
    • Yibo Hou
    • Jiayun Hu
    • Yongshen He
    • Jiajie Chen
    • Kunmei Ji
  • View Affiliations

  • Published online on: March 18, 2019     https://doi.org/10.3892/mmr.2019.10050
  • Pages: 3497-3504
  • Copyright: © Zhang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The detection of allergen‑specific immunoglobulin (Ig)E is an important method for the diagnosis of IgE‑mediated allergic diseases. The sensitivity of the indirect IgE‑ELISA method against allergen extracts is limited by interference from high IgG titers and low quantities of effectual allergen components in extracts. To overcome these limitations, a novel capture IgE‑ELISA based on a recombinant Der f 1/Der f 2 fusion protein (rDer f 1/2) was developed to enhance the sensitivity to IgEs that bind allergens from the house dust mite (HDM) species Dermatophagoides farina. pET28‑Der f 1/2 was constructed and expressed in Escherichia coli BL21 (DE3) pLysS. The purified fusion protein was evaluated by IgE western blotting, IgE dot blotting and indirect IgE‑ELISA. Capture‑ELISA was performed by coating wells with omalizumab and incubating in series with sera, biotinylated Der f 1/2, horseradish peroxidase‑conjugated streptavidin and 3,3,5,5‑tetramethylbenzidine. The relative sensitivities of indirect‑ELISA and capture‑ELISA for HDM allergen‑specific IgE binding were determined; sera from non‑allergic individuals were used as the control group. rDer f 1/2 was expressed in the form of inclusion bodies comprising refolded protein, which were then purified. It exhibited increased IgE‑specific binding (24/28, 85.8%) than rDer f 1 (21/28, 75.0%) or rDer f 2 (22/28, 78.6%) with HDM‑allergic sera. Furthermore, in a random sample of HDM‑allergic sera (n=71), capture‑ELISA (71/71, 100%) was more sensitive than indirect‑ELISA (68/71, 95.8%) for the detection of HDM‑specific IgEs (P<0.01), indicating that this novel method may be useful for the diagnosis of HDM allergy.
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May-2019
Volume 19 Issue 5

Print ISSN: 1791-2997
Online ISSN:1791-3004

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Spandidos Publications style
Zhang Z, Cai Z, Hou Y, Hu J, He Y, Chen J and Ji K: Enhanced sensitivity of capture IgE‑ELISA based on a recombinant Der f 1/2 fusion protein for the detection of IgE antibodies targeting house dust mite allergens. Mol Med Rep 19: 3497-3504, 2019
APA
Zhang, Z., Cai, Z., Hou, Y., Hu, J., He, Y., Chen, J., & Ji, K. (2019). Enhanced sensitivity of capture IgE‑ELISA based on a recombinant Der f 1/2 fusion protein for the detection of IgE antibodies targeting house dust mite allergens. Molecular Medicine Reports, 19, 3497-3504. https://doi.org/10.3892/mmr.2019.10050
MLA
Zhang, Z., Cai, Z., Hou, Y., Hu, J., He, Y., Chen, J., Ji, K."Enhanced sensitivity of capture IgE‑ELISA based on a recombinant Der f 1/2 fusion protein for the detection of IgE antibodies targeting house dust mite allergens". Molecular Medicine Reports 19.5 (2019): 3497-3504.
Chicago
Zhang, Z., Cai, Z., Hou, Y., Hu, J., He, Y., Chen, J., Ji, K."Enhanced sensitivity of capture IgE‑ELISA based on a recombinant Der f 1/2 fusion protein for the detection of IgE antibodies targeting house dust mite allergens". Molecular Medicine Reports 19, no. 5 (2019): 3497-3504. https://doi.org/10.3892/mmr.2019.10050