Methods for detecting biomolecules in a sample

A high-throughput method for detecting at least two different target biomolecules in a sample, which comprises the steps of:

• (a) mixing at least two different fluorophore-labeled peptide probes with a specimen that contains said at least two different target biomolecules of respective probes, each said respective peptide probe only specifically binds to its respective said target biomolecule;
• (b) adding dextran-coated charcoal to the reaction mixture to capture all of the unbound fluorophore-labeled peptide probes, thereby adsorbing said unbound fluorophore-labeled peptide probes and causing said unbound fluorophore-labeled peptide probes to not exhibit fluorescent properties;
• (c) precipitating the dextran-coated charcoal by centrifugation or magnetic sedimentation so that unbound fluorophore-labeled peptide probes are separated from a supernatant;
• and
• (d) detecting the signals of the fluorophore in the supernatant with respective detection parameters while said unbound fluorophore-labeled peptide probes and said supernatant are still in a same container;
• wherein the at least two target biomolecules are large molecules that cannot be adsorbed by the dextran-coated charcoal and have a molecular weight of more than about 10 KD, and the respective probes are small molecules containing no more than 18 amino acids which can be adsorbed by the dextran-coated charcoal;
• and wherein after charcoal precipitation in step (c), it is unnecessary to transfer biomolecule-peptide probe complex to a new container for detection of fluorescence.