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Updated: Aug 9, 2026

Multiplexed Fluorometric ImmunoAssay Testing Methodology and Troubleshooting
Published on: December 12, 2011
Immunoassay for IgG rheumatoid factor with a murine monoclonal anti-Fd antibody
Conventional radioimmunoassays for IgG rheumatoid factors (RF) detect the binding of IgG RF (or their F(ab')2 fragments) to solid-phase human IgG Fc fragments or rabbit IgG. Binding is detected with a radiolabeled antibody that is IgG-specific but is nonreactive with human Fc (i.e., an alpha-Fd reagent) or with rabbit IgG. Anti-Fd reagents are quite laborious to produce. We developed a murine monoclonal alpha-Fd antibody, confirmed its specificity, and compared it with conventional rabbit alpha-Fd in the IgG RF radioimmunoassay. We also adapted the assay to an enzyme-linked assay with both human Fc and rabbit IgG as antigen. We compared the four assay methods with each other and examined their relationship to certain clinical and laboratory features of rheumatoid arthritis.
Conventional radioimmunoassays for IgG rheumatoid factors (RF) detect the binding of IgG RF (or their F(ab')2 fragments) to solid-phase human IgG Fc fragments or rabbit IgG. Binding is detected with a radiolabeled antibody that is IgG-specific but is nonreactive with human Fc (i.e., an alpha-Fd reagent) or with rabbit IgG. Anti-Fd reagents are quite laborious to produce. We developed a murine monoclonal alpha-Fd antibody, confirmed its specificity, and compared it with conventional rabbit alpha-Fd in the IgG RF radioimmunoassay. We also adapted the assay to an enzyme-linked assay with both human Fc and rabbit IgG as antigen. We compared the four assay methods with each other and examined their relationship to certain clinical and laboratory features of rheumatoid arthritis.
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