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Engineered chimeric myeloperoxidase (MPO) antibodies as candidate reference materials for MPO-ANCA detection
Mingming Zhang1, Hong Chen1, Fangming Cheng2
1Key Laboratory of Developmental Genes and Human Diseases, School of Life Science and Technology, Southeast University, Nanjing, Jiangsu 210088, China.
Journal of Immunological Methods
|March 11, 2026
Summary
Researchers developed chimeric antibodies to standardize anti-neutrophil cytoplasmic antibodies (ANCA) testing for myeloperoxidase (MPO). These antibodies show consistent reactivity and can serve as reference materials for improved MPO-ANCA assay harmonization and evaluation.
Area of Science:
- Immunology
- Biotechnology
- Clinical Diagnostics
Background:
- Standardization of anti-neutrophil cytoplasmic antibodies (ANCA) targeting myeloperoxidase (MPO) is crucial for diagnosing microscopic polyangiitis (MPA).
- Current MPO-ANCA testing faces challenges due to method diversity and lack of quantitative reference standards.
Purpose of the Study:
- To develop and characterize human-mouse chimeric monoclonal antibodies targeting conformational MPO epitopes.
- To establish candidate quantitative reference materials for MPO-ANCA detection immunoassays.
Main Methods:
- Screened hybridoma-derived murine anti-MPO clones for native MPO reactivity.
- Engineered high-affinity clones (B61, B62) with human IgG1 constant domains.
- Assessed binding specificity and diagnostic utility via indirect immunofluorescence (IIF), line immunoassay (LIA), ELISA, and chemiluminescence immunoassay (CLIA).
Main Results:
- Chimeric B61 and B62 antibodies exclusively bound native MPO, showing no reactivity with denatured MPO.
- Antibodies produced characteristic p-ANCA staining in IIF and confirmed specificity in LIA.
- Quantitative ELISA and CLIA yielded standard curves (R² > 0.99); competition assays showed partial epitope overlap with patient-derived MPO-ANCAs.
Conclusions:
- Engineered chimeric antibodies exhibit consistent native MPO reactivity and compatibility with multiple MPO-ANCA assays.
- Proposed as candidate reference materials for assay harmonization and performance evaluation.
- May support titer assessments or serve as alternatives to MPO-positive serum controls in research/preliminary clinical settings.
