Patent ID: 11965884
Assignee: ALEXION PHARMACEUTICALS, INC.
Field: Measurement (Instruments)
Classification: CPC G | IPC G

Claim 0:
1. A method of quantitating unbound human C5 complement protein (C5) in a sample obtained from a human patient before and after treatment with an anti-C5 antibody, wherein the method comprises:
performing separate assays on the pre-treatment sample and post-treatment sample, wherein each of the assays respectively comprises:
a. binding biotinylated anti-C5 capture antibody to streptavidin coated particles; wherein said biotinylated anti-C5 capture antibody is added by capillary action to a compact disc (CD) comprising columns with the streptavidin coated particles; wherein said CD is subjected to centrifugal force inside an instrument, thus driving the biotinylated anti-C5 capture antibody to the streptavidin coated particles in the columns;
b. capturing the unbound C5 in the pre-treatment sample or the post-treatment sample; wherein the pre-treatment sample or the post-treatment sample is added to the CD by capillary action; wherein said CD is subjected to centrifugal force inside the instrument, thus driving the pre-treatment sample or the post-treatment sample to the biotinylated anti-C5 capture antibody bound to the streptavidin coated particles in the columns;
c. detecting the captured C5; wherein an Alexa Fluor labeled anti-C5 detection antibody is added to the CD by capillary action, wherein said anti-C5 detection antibody binds C5 at a different epitope from the epitope bound by the capture antibody;
wherein said CD is subjected to centrifugal force inside the instrument, thus driving the detection antibody to the C5 bound to the capture antibody bound to the streptavidin coated particles in the columns; and
d. quantitating the captured C5 using laser-induced fluorescence detection, wherein the pre-treatment sample is diluted at least 1:20 and post-treatment sample is diluted about 1:2.