Controls for implementing multiplex analysis methods

A multiplex analysis method for detecting at least n analytes in at least one sample, n being an integer greater than or equal to 2, said method comprising at least steps a), c) and e) or at least steps a), b), c) and d) as follows:

• a) providing at least one solid support,
• b) placing, in the presence of the spots of said compartment:
• l detection ligands of p analytes to be detected and at least one additive, l being greater than or equal to p,
• c) placing a sample to be analyzed in the presence of the spots of said compartment,
• d) placing, in the presence of the spots of said compartment:
• l′ detection ligands of m analytes to be detected, and, at least one detection ligand of said additive(s), l′ being greater than or equal to m, and
• e) placing, in the presence of the spots of said compartment:
• at least one detection ligand of a control compound and l″ detection ligands of y analytes to be detected, l″ being greater than or equal to y,
• l, l′, l″, m, p and y being integers greater than or equal to 0 and the sum m+p+y being greater than or equal to 1;
• said solid support comprising at least one compartment,
• said compartment comprising a plurality of spots, each spot being deposited at a separate location within said at least one compartment,
• said plurality of spots comprising two or more control spots deposited upon said solid support within said at least one compartment and at least two detection spots comprising a ligand specific for an analyte in a sample,
• and said at least two control spots being selected from the group consisting of a spot for controlling the deposition of a sample, a spot for controlling the deposition of an analyte detection ligand and a spot for controlling the deposition of a reporter, wherein:
• a) the spot for controlling the deposition of the sample comprises at least one control capture ligand specific for a control compound naturally present in the sample to be analyzed, said control capture ligand being an antibody, antigen, peptide, carbohydrate, lipid, or nucleic acid;
• b) the spot for controlling the deposition of the detection ligand for the analyte comprises at least one additive capture ligand specific for an additive that is added to a sample and said additive is not naturally present in the sample or derived from any compound present in the sample, said at least one additive capture ligand being an antibody, antigen, peptide, carbohydrate, lipid, or nucleic acid;
• c) the spot for controlling the deposition of the reporter comprises a direct marker, an indirect marker or a carrier molecule coupled to an indirect marker, wherein the reporter specifically interacts with the indirect marker or the indirect marker coupled to the carrier molecule, optionally, in presence of a substrate, to produce a detectable signal,
• said direct marker being selected from a radioisotope, a fluorochrome, a lanthanide, a luminescent compound, a transition metal, and colored, fluorescent or luminescent nanoparticles,
• said indirect marker being an enzyme, biotin, avidin, streptavidin, neutravidin, a hapten, an antigen or an antibody,
• said carrier being a polypeptide, functionalized polymer, a copolymer, or an antibody, and
• said reporter being an enzymatic substrate for an enzyme, an enzyme for a luminescent compound, and avidin, streptavidin, or neutravidin coupled to a direct or indirect marker for biotin, and
• said ligands for analyte detection are antibodies, antigens, peptides, carbohydrates, lipids, or nucleic acids.