Related Experiment Videos
ELISA for evaluating the incorporation of plasma derived complement split-products C3b/iC3b into solid-phase immune
E Zimmermann-Nielsen1, S E Svehag, O Thorlacius-Ussing
1Department of Surgical Gastroenterology, Aalborg Hospital, Hobrovej 18-22, Postbox 365, 9100, Aalborg, Denmark. otu@aas.nja.dk
Journal of Immunological Methods
|February 28, 2001
Summary
This study introduces a new ELISA assay to measure complement activation capacity. The assay revealed no significant differences in complement activation pathways based on age or gender in healthy donors.
Area of Science:
- Immunology
- Biochemistry
Background:
- Complement system activation produces split products like C3b/iC3b.
- Assessing complement activation capacity is crucial for understanding immune responses.
Purpose of the Study:
- To describe a novel ELISA for measuring complement (C) activation capacity.
- To evaluate the assay's performance in healthy individuals and patients with systemic lupus erythematosus (SLE).
Main Methods:
- Developed an ELISA using solid-phase immune complexes to detect deposited C3b/iC3b.
- Coated plates with BSA and anti-BSA, added plasma, and detected C3b/iC3b with biotinylated anti-C3c antibodies and avidin-alkaline phosphatase.
- Compared ELISA results with a standard hemolytic complement CH(50) assay in SLE patients.
Main Results:
- The ELISA demonstrated high reproducibility (CV <5.7%) and found no age or gender-related differences in alternative pathway (AP) or classical pathway (CP) activation capacity in 250 healthy donors.
- A weak correlation was observed between the ELISA and CH(50) assay for both pathways in SLE patients.
- The CP capacity was significantly reduced in SLE patients compared to healthy controls (P<0.0001).
Conclusions:
- The developed ELISA is a reliable method for assessing complement activation capacity.
- While not correlating with ACR criteria for SLE, the assay detected reduced classical pathway function in SLE patients, suggesting its potential clinical utility.