Method for simultaneous analysis of multiple biological reactions or changes in in vivo conditions

A method for analyzing enzyme activity comprising:

• mixing a mixture including at least one activity measurement molecule with a test sample, wherein said activity measurement molecule comprises a biomolecule covalently bonded to a quantum dot and a fluorescent protein, wherein said biomolecule is selected from the group consisting of a nucleic acid, a protein, a peptide, an amino acid, a coenzyme, a saccharide, a poly saccharide, a lipid, derivatives thereof, and complexes thereof but excludes double stranded DNA, wherein said biomolecule is a substrate of an enzyme reaction and said test sample includes at least one enzyme used as a measurement target;
• incubating the mixture under conditions in which said biomolecule reacts with said enzyme;
• exciting each said quantum dot with radiation of a wavelength suitable for said quantum dot;
• and
• detecting a change in fluorescence intensity obtained from at least one of said quantum dot and said fluorescent protein as a result of said enzyme reaction to measure said activity of said enzyme.

A method for analyzing enzyme activity comprising:

• mixing a mixture including at least one activity measurement molecule with a test sample, wherein said activity measurement molecule comprises a biomolecule covalently bonded to a quantum dot and a fluorescent protein, wherein said biomolecule is selected from the group consisting of a nucleic acid, a protein, a peptide, an amino acid, a coenzyme, a saccharide, a poly saccharide, a lipid, derivatives thereof, and complexes thereof but excludes double stranded DNA, wherein said biomolecule is designed to have a portion that is susceptible to an enzyme reaction and said test sample includes at least one enzyme used as a measurement target;
• incubating the mixture under conditions in which said enzyme reacts with said enzyme susceptible portion of said biomolecule;
• exciting said quantum dot with radiation of a wavelength suitable for fluorescent emission from said quantum dot;
• wherein said quantum dot is a FRET donor and said fluorescent protein is positioned in proximity to said quantum dot as a FRET acceptor such that excitement of said fluorescent protein acceptor occurs by fluorescent resonance energy transfer as said fluorescent emission from said excited quantum dot donor, and
• detecting a change in fluorescence intensity of said radiation emitted from at least one of said quantum dot and said fluorescent protein as a result of said enzyme reaction to measure said activity of said enzyme.

A method for analyzing enzyme activity comprising:

• mixing a mixture including at least one activity measurement molecule with a test sample, wherein said activity measurement molecule comprises a biomolecule covalently bonded to a quantum dot and a fluorescent protein, wherein said biomolecule is selected from the group consisting of a nucleic acid, a protein, a peptide, an amino acid, a coenzyme, a saccharide, a poly saccharide, a lipid, derivatives thereof, and complexes thereof but excludes double stranded DNA, wherein said biomolecule is designed to have a portion that is susceptible to an enzymatic reaction, said mixture comprising at least one other activity measurement molecule with a different biomolecule covalently bonded to a different quantum dot and a different fluorescent molecule, wherein said different biomolecule is selected from the group consisting of a nucleic acid, a protein, a peptide, an amino acid, a coenzyme, a saccharide, a poly saccharide, a lipid, derivatives thereof, and complexes thereof but excludes double stranded DNA, wherein said different biomolecule is a substrate of an enzyme reaction that is different from said enzyme reaction for which said biomolecule of said activity measurement molecule is said substrate, said different quantum dot having a different fluorescent property than said quantum dot of said activity measurement molecule, and said test sample includes as measurement targets at least two different enzymes in said mixture;
• incubating the mixture under conditions in which one of said enzymes reacts with said biomolecule of said activity measurement molecule and another of said enzymes reacts with said different biomolecule;
• applying radiation of the same wavelength to said quantum dot of said activity measurement molecule and said different quantum dot, said radiation having a wavelength suitable for fluorescent emission from said quantum dot, said fluorescent protein, said different quantum dot and said fluorescent molecule,
• emitting radiation of different wavelengths from said quantum dot of said activity measurement molecule and said different quantum dot, and
• detecting a change in fluorescence intensity of said radiation emitted from at least one of said quantum dot, said different quantum dot, said fluorescent protein and said fluorescent molecule, as a result of said enzyme reactions to measure said activity of said enzymes.