Systematic reference sample generation for multiplexed serological assays

Quality controls of serological assays have to contain defined amounts of human antibodies specific for the targeted antigen. A prevailing issue for array-based antigen assays is that dozens of antigens are targeted within the same assay. Commonly different patient sera are combined and optimal pools are empirically identified. Here, we report a mathematical approach how an optimal sample pool composition can be systematically calculated and accurately compiled. The approach was used to compose suitable quality controls for a 71 plex Tuberculosis antigen bead array using a limited number of positive human sera.