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Identification and Characterization of Immunogenic RNA Species in HDM Allergens that Modulate Eosinophilic Lung Inflammation
Published on: May 30, 2020
Development and application of novel immunoassays for eosinophil granule major basic proteins to evaluate
Diane L Squillace1, James L Checkel1, Ayalew Tefferi2
1Division of Allergic Diseases, Department of Medicine, Mayo Clinic, Rochester, MN, United States of America.
Background:
During eosinophil differentiation, the granule eosinophil major basic protein 1 (eMBP1) is synthesized as a 32-kDa precursor form, referred to as proMBP1, which is processed into the 14-kDa mature form of eMBP1. The prevalence of these two forms of MBP1 in most pathological conditions remains unknown.
Objective:
To develop the immunoassays that differentiate mature eMBP1 and proMBP1 and apply them to analyze their levels in biological fluids from patients with eosinophilia and hematologic disorders.
Methods:
We produced a series of monoclonal antibodies and selected pairs capable of discriminating between the two molecular forms of eMBP1. Radioimmunoassay (RIA) was performed to simultaneously quantitate the levels of mature eMBP1 and proMBP1 in secretions from patients with chronic rhinosinusitis (CRS) and sera from patients with hypereosinophilic syndrome (HES) and other myeloproliferative disorders.
Results:
The novel immunoassays possessed less than 1% crossreactivity between mature eMBP1 and proMBP1. Mature eMBP1, but not proMBP1, was found in nasal secretions of CRS patients. In contrast, elevated serum levels of mature eMBP1 and proMBP1 were observed in approximately 60% and 90% of HES patients, respectively, with proMBP1 present in greater quantities than mature eMBP1. Patients with several myeloproliferative disorders also showed high serum levels of proMBP1 while mature eMBP1 remained at basal levels.
Conclusion:
The novel immunoassays successfully differentiated mature eMBP1 and proMBP1 in human biological fluids. Further studies addressing the clinical correlates of these assays will help to develop biomarkers to diagnose and monitor patients with eosinophilia and myeloproliferative disorders.
Insights
New immunoassays can distinguish between mature eosinophil major basic protein 1 (eMBP1) and its precursor, proMBP1. Elevated proMBP1 levels were found in hypereosinophilic syndrome and myeloproliferative disorders, suggesting potential diagnostic biomarkers.
Area of Science:
- Immunology
- Hematology
- Biochemistry
Background:
- Eosinophil differentiation involves processing of eosinophil major basic protein 1 (eMBP1) from a precursor (proMBP1) to a mature form.
- The distinct prevalence of mature eMBP1 and proMBP1 in pathological conditions is largely unknown.
Purpose of the Study:
- To develop novel immunoassays capable of differentiating mature eMBP1 from proMBP1.
- To quantify and analyze the levels of both eMBP1 forms in biological fluids from patients with eosinophilia and hematologic disorders.
Main Methods:
- Production and selection of monoclonal antibodies for specific detection of mature eMBP1 and proMBP1.
- Simultaneous quantification of mature eMBP1 and proMBP1 using radioimmunoassay (RIA) in patient samples.
Main Results:
- Novel immunoassays demonstrated high specificity with less than 1% cross-reactivity.
- Mature eMBP1 was detected in nasal secretions of chronic rhinosinusitis (CRS) patients, while proMBP1 was not.
- Elevated serum levels of both mature eMBP1 and proMBP1 were observed in hypereosinophilic syndrome (HES) patients, with proMBP1 being more abundant. Myeloproliferative disorders showed high proMBP1 but basal mature eMBP1.
Conclusions:
- Developed immunoassays effectively differentiate mature eMBP1 and proMBP1 in human biological fluids.
- These assays hold potential for developing biomarkers to diagnose and monitor patients with eosinophilia and myeloproliferative disorders.
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