Abstract
Flow cytometric methods are commonly used to analyze white blood cell surface antigen expression. We developed a flow cytometric method to detect red blood cell (RBC)-bound immunoglobulin (Ig)G, IgA, and IgM. RBCs were washed; incubated with fluorescein isothiocyanate (FITC)-conjugated anti-IgG, -IgA, or -IgM; washed; and analyzed on the flow cytometer. The method was optimized by determining the dilution of FITC-conjugated anti-IgG, -IgA, and -IgM providing the greatest amount of fluorescence when tested with Ig-coated RBCs and the least amount of fluorescence when tested with naive RBCs. Tannic acid was used to prepare Ig-coated RBCs. Cross-reactivity of FITC-conjugated anti-IgG, -IgA, and -IgM with Ig-coated RBCs was evaluated, and a reference range was established. Use of this method may assist in clinical evaluation of patients who present with hemolysis and a negative direct antiglobulin test.
Original language | English (US) |
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Pages (from-to) | 161-169 |
Number of pages | 9 |
Journal | Immunohematology |
Volume | 32 |
Issue number | 4 |
State | Published - 2016 |
Keywords
- Autoimmune hemolytic anemia
- Direct antiglobulin test
- Flow cytometry
- Immunoglobulins
- Red blood cell
- Tannic acid
ASJC Scopus subject areas
- Immunology and Allergy
- Hematology