| 摘要 |
Methods and devices for evaluating a sample, e.g., a plasma sample, from a subject, for detecting a target red blood cell protein or antibody are disclosed. In one embodiment, optimized antibody screening methods and devices significantly reduce the level of non-specific binding to a surface (e.g., a test surface bound with a red blood cell (rbcm) preparation), thus allowing for more efficient detection and reduced test time. In one embodiment, the optimized antibody screening method includes an immunoglobulin G (IgG) binding moiety that binds selectively and specifically to the plasma IgG present relative to the binding to the lysed rbcm preparation. In another embodiment, an optimized antibody screening method is disclosed whereby non-specific binding caused by lysed red blood cell membrane preparations can be reduced by an agent that specifically cleaves a human IgG in the hinge region. In other embodiments, the invention provides methods and devices for target capturing that include a substantially planar surface, optionally having an optimized angle, for capture. Alternative solid phase geometries for capture are disclosed. Optimized methods for cell deposition are also disclosed. Thus, optimized methods, devices, kits, assays for evaluating a sample are disclosed. |
| 主权项 |
1. A method of detecting an antibody of a G isotype (IgG antibody) against a red blood cell (RBC) antigen in a sample from a subject, comprising:
(a) contacting a first red blood cell membrane preparation (a rbcm preparation) comprising a first RBC antigen, with the sample from said subject, under conditions sufficient for the formation of an immune complex between said first RBC antigen and an anti-first-RBC antigen IgG antibody in said sample, wherein said first rbcm preparation is immobilized on a surface or substrate; and (b) contacting a detection reagent with the immune complex of (a) under conditions sufficient for the formation of an immune complex between said detection reagent and the anti-first-RBC antigen IgG antibody in said sample, said detection reagent comprising an IgG-specific binding moiety, thereby detecting an anti-RBC antigen IgG antibody in a sample. |
