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Specificity of three anti-complement factor 3 monoclonal antibodies
S Henwick1, S V Hetherington, M K Hostetter
1Department of Infectious Diseases, St. Jude Children's Research Hospital, Memphis, TN 38101-0318.
Journal of Immunological Methods
|August 30, 1992
Summary
Monoclonal antibodies targeting complement component 3 (C3) fragments are crucial for identifying C3 cleavage products. Researchers found one antibody lost specificity on non-erythrocyte surfaces, highlighting the need for rigorous testing.
Area of Science:
- Immunology
- Biochemistry
Background:
- Monoclonal antibodies (mAbs) are vital tools for detecting complement component 3 (C3) fragments.
- Accurate identification of C3 cleavage products bound to various surfaces is essential in immunological studies.
Purpose of the Study:
- To evaluate the specificity of three commercially available monoclonal antibodies against C3 fragments.
- To assess the reliability of these antibodies across different assay formats, particularly concerning erythrocyte-bound versus non-erythrocyte-bound C3 fragments.
Main Methods:
- Western immunoblot analysis
- Enzyme-linked immunosorbent assay (ELISA)
- Quantitative flow cytometry
Main Results:
- Two monoclonal antibodies specific for C3d and C3c epitopes retained their specificity across all tested assays.
- A third monoclonal antibody, initially selected for C3bi specificity, exhibited cross-reactivity with C3g on non-erythrocyte surfaces.
- Erythrocyte surface proteins may influence the binding specificity of anti-C3 antibodies.
Conclusions:
- The specificity of monoclonal antibodies for C3 fragments must be validated using non-erythrocyte-based assays.
- Quantitative flow cytometry offers a valuable method for enumerating particle-bound C3 fragments.