Complement assays and uses thereof

摘要

The present invention provides methods for assessing complement activation and methods for assessing the ability of an agent or condition of interest to modulate complement activation. The present invention provides methods for assessing whether a subject has or is at increased risk of developing a complement-mediated disorder, e.g., age-related macular degeneration (AMD). Also provided are kits containing materials useful for performing the methods.

主权项

1. A method of assessing complement activation in a sample, the method either comprising steps of:
(a) capturing intact C3 present in the sample or in a sub-sample thereof using an antibody that specifically binds to at least one epitope that is present in intact C3; (b) quantifying intact C3 captured in step (a) using an antibody that specifically binds to at least one epitope of intact C3 and has a detectable label directly or indirectly linked thereto; (c) capturing iC3b present in the sample or in a sub-sample thereof using an antibody that specifically binds to a neoepitope of iC3b; (d) quantifying iC3b captured in step (c) using an antibody that specifically binds to at least one epitope of iC3b and has a detectable label directly or indirectly linked thereto; and (e) calculating the ratio of iC3b to intact C3 using the results of (b) and (d) or comprising steps of: (i) capturing intact C3 present in the sample or in a sub-sample thereof using an antibody that specifically binds to at least one epitope that is present in intact C3; (ii) quantifying intact C3 captured in step (i) using an antibody that specifically binds to at least one epitope of intact C3 and has a detectable label directly or indirectly linked thereto; (iii) capturing total C3 present in the sample or in a sub-sample thereof using an antibody that binds to total C3, wherein the antibody specifically binds to at least one epitope that is present in total C3; (iv) quantifying total C3 captured in step (iii) using an antibody that specifically binds to at least one epitope of C3c or C3d and has a detectable label directly or indirectly linked thereto; and (v) calculating the relative concentration of intact C3 and total C3 in the sample using the results of (ii) and (iv).