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Updated: Aug 8, 2026
High-throughput Flow Cytometry Cell-based Assay to Detect Antibodies to N-Methyl-D-aspartate Receptor or Dopamine-2 Receptor in Human Serum
Published on: November 23, 2013
Flow cytometric analysis of 68 monoclonal anti-D reagents
Insights
Monoclonal IgG anti-D antibodies showed varied binding to D variant red blood cells. Some D variants lack specific epitopes, impacting antibody recognition and potentially transfusion compatibility.
Area of Science:
- Immunology
- Hematology
- Blood group serology
Background:
- The RhD antigen is crucial for blood transfusion compatibility.
- D variants present challenges in accurately determining RhD status.
- Monoclonal antibodies are key tools in blood group antigen detection.
Purpose of the Study:
- To quantitatively and qualitatively assess the binding of monoclonal IgG anti-D antibodies to D variant red blood cells.
- To investigate epitope expression on different D variants.
- To understand the implications for RhD typing.
Main Methods:
- Utilized flow cytometry and indirect immunofluorescence assays.
- Tested 68 monoclonal IgG anti-D antibodies.
- Evaluated binding to normal RhD positive red cells and various D variant red cells (D category III, VI, VII, R1VIr, R2VIr, R1VIIr, DFR).
Main Results:
- Binding varied significantly (up to twenty-fold) between different monoclonal antibodies and D variants.
- D category III cells showed the lowest binding levels compared to controls.
- Certain D variants (R1VIr, R2VIr, R1VIIr, DFR) failed to react with specific monoclonal antibodies, suggesting missing epitopes.
Conclusions:
- Monoclonal anti-D antibodies exhibit differential binding to D variants.
- The presence or absence of specific epitopes on D variants influences antibody binding.
- These findings support the concept of missing epitopes in certain D variants, impacting serological detection.
Abstract:
68 monoclonal IgG anti-D were assessed for quantitative and qualitative binding to D variant red cells using a flow cytometric indirect immunofluorescence test. One antibody failed to bind to normal RhD pos. cells, but reacted weakly with D cat VI and D Cat VII cells. Quantitatively, binding varied approximately twenty-fold between different MAbs and different D variants. D cat III cells gave the lowest level of binding when compared with the D pos. controls. Qualitatively, R1VIr, R2VIr, R1VIIr and DFR samples failed to react with various MAbs regardless of the levels of binding of these MAbs to the D pos. controls, thus supporting the idea of missing epitopes in such variants.
