Patent ID: 11913960
Assignee: THE UNIVERSITY OF TOLEDO
Field: Measurement (Instruments)
Classification: CPC G | IPC G

Claim 0:
1. A method for conducting an enzyme linked immunosorbent assay (ELISA) for detecting the activity of properdin, the method comprising:
i) coating a surface with a non-inhibitory anti-properdin antibody, and incubating for a first desired time and at a first desired temperature;
ii) adding a blocking agent to prevent non-specific binding and to block any remaining active sites, and incubating for a second desired time and at a second desired temperature;
iii) adding a normal human serum sample that contains properdin to the surface of step ii), wherein the normal human serum that contains properdin is added in ethylenediaminetetraacetic acid (EDTA), and incubating for a third desired time and at a third desired temperature;
iv) adding a properdin-depleted serum sample to the surface of step iii), wherein the properdin-depleted serum sample provides complement proteins including C3(H2O), Factor B, and Factor D, and,
wherein C3(H2O) and Factor B bind to properdin, and wherein Factor D cleaves Factor B to form C3(H2O)Bb that, in turn, cleaves C3 and deposits C3b covalently on the blocking agent on the surface; and
v) adding an anti-C3b antibody to detect the C3b on the blocking agent on the surface.