Patent ID: 8715951
Filing Date: 2014-05-06
Classification: C07K,G01N,Y10S,Y10T

Abstract:
1. A method for determining the level of a FVIII protein or a fragment thereof in a sample, comprising: (a) contacting the sample with a first antibody directed to a FVIII antigen, said first antibody being attached to a first bead having at least one first characteristic, wherein the FVIII protein or fragment thereof comprising said FVIII antigen binds to said first antibody being attached to the first bead, thereby forming a capture antibody, and forms a complex therewith; (b) contacting the sample with a control antibody not directed to a FVIII antigen, said control antibody being attached to a second bead having at least one second characteristic distinguishable from the first characteristic of the first bead, wherein one or more proteins in the sample may form non-specific complexes with the control antibody; (c) contacting the complexes formed in steps (a) and (b) with a second antibody directed to the FVIII antigen, said second antibody being labeled with a marker, to form a complex which includes the antibody of step (a) or (b), the FVIII protein or fragment thereof comprising said FVIII antigen, and the second antibody; and (d) subtracting the quantity of the second labeled antibody in complexed associated with the second beads attached to the control antibody from the quantity of the second labeled antibody in complexes associated with the first beads attached to the first antibody directed against the FVIII antigen, thereby removing spurious non-specific binding and more accurately determining the level of the FVIII protein or fragment thereof comprising said antigen in the sample.