Porphyrinic compounds for use in flow cytometry

1. In a method of detecting a target compound, cell or particle, wherein said target is labelled with a detectable luminescent compound, the improvement comprising utilizing as said detectable luminescent compound a compound of the formula A-A′-Z—B′—B, wherein:

A is a targeting group that specifically binds said detectable luminescent compound to said target compound, cell or particle, said targeting group selected from the group consisting of protein, peptide and antibody;

A′ is a linking group or covalent bond;

B′ is a linking group or covalent bond;

B is a water-soluble group containing at least one ionic or polar group selected from the group consisting of hydroxy, alkoxy, carboxy, nitro, cyano, amino, amido, ureido, sulfonamido, sulfinyl, sulfhydryl, silyl, S-sulfonamido, N-sulfonamido, C-carboxy, O-carboxy, C-amido, N-amido, sulfonyl, phosphono, morpholino, piperazinyl, tetrazolo, alcohol, thiol, polyethylene glycol, polyol, sulfonamide, carboxamide, hydrazide, N-hydroxycarboxamide, urea, metal chelates, carboxylate, sulfonate, phosphate, amine N-oxide, ammonium, uronic acid, carboxylic acid, sulfonic acid, amine, guanidinium, phosphoric acid, phosphonic acid, phosphatidyl choline, phosphonium, borate sulfate, and combinations thereof; and

Z is chlorin.

2. The method of claim 1 , wherein said detecting is carried out by flow cytometry.

3. The method of claim 1 , wherein said detectable luminescent compound is excited by light at an excitation wavelength band and detected by emitted light at an emission wavelength band;

and wherein said emission wavelength band has a full width at half maximum peak of not more than 100 nanometers.

4. The method of claim 3 , wherein said emission wavelength band has a full width at half maximum peak of not more than 30 nanometers.

5. The method of claim 3 , wherein

said excitation wavelength band has a peak between 350 and 900 nanometers; and

said emission wavelength band has a peak between 550 and 1000 nanometers.

6. The method of claim 3 , wherein:

said porphyrinic macrocycle is a chlorin;

said excitation wavelength band has a peak between 600 and 740 nanometers; and

said emission wavelength band has a peak between 610 and 800 nanometers.

7. The method of claim 3 , wherein:

said porphyrinic macrocycle is a chlorin;

said excitation wavelength band has a peak between 350 and 450 nanometers;

said emission wavelength band has a peak between 600 and 800 nanometers.

8. The method of claim 1 , wherein said detectable luminescent compound is excited by light at an excitation wavelength band and detected by emitted light at an emission wavelength band;

wherein said excitation wavelength band and said emission wavelength band each have peaks that are separated from one another by at least 100 nanometers;

and wherein said emission wavelength band has a full width at half maximum peak of not more than 100 nanometers.

9. The method of claim 1 , wherein said detectable luminescent compound is excited by light at an excitation wavelength band and detected by emitted light at an emission wavelength band;

wherein said excitation wavelength band and said emission wavelength band each have peaks that are separated from one another by from 10 to 100 nanometers;

and wherein said emission wavelength band has a full width at half maximum peak of not more than 25 nanometers.

10. In a method of detecting and distinguishing first and second target compounds, cells or particles, wherein said first target is labelled with a first detectable compound and said second target is labeled with a second detectable compound, the improvement comprising:

utilizing a compound comprising a first porphyrinic macrocycle as said first detectable compound and a compound comprising a second porphyrinic macrocycle as said second detectable compound;

wherein each of said first and second detectable compounds are excited at an excitation wavelength band;

wherein said first detectable compound comprises a compound of the formula A 1 -A 1′ -Z 1 —B 1 ′—B 1 , wherein:

A 1 is a targeting group that specifically binds said detectable compound to said target compound, cell or particle, said targeting group selected from the group consisting of protein, peptide, and antibody;

A 1 ′ is a linking group or covalent bond;

B 1 ′ is a linking group or covalent bond;

B 1 is a water-soluble group containing at least one ionic or polar group selected from the group consisting of hydroxy, alkoxy, carboxy, nitro, cyano, amino, amido, ureido, sulfonamido, sulfinyl, sulfhydryl, silyl, S-sulfonamido, N-sulfonamido, C-carboxy, O-carboxy, C-amido, N-amido, sulfonyl, phosphono, morpholino, piperazinyl, tetrazolo, alcohol, thiol, polyethylene glycol, polyol, sulfonamide, carboxamide, hydrazide, N-hydroxycarboxamide, urea, metal chelates, carboxylate, sulfonate, phosphate, amine N-oxide, ammonium, uronic acid, carboxylic acid, sulfonic acid, amine, guanidinium, phosphoric acid, phosphonic acid, phosphatidyl choline, phosphonium, borate, sulfate, and combinations thereof; and

Z 1 is chlorin; and

said second detectable compound comprises a compound of formula A 2 -A 2′ -Z 2 —B 2′ —B 2 , wherein:

A 2 is a targeting group that specifically binds said detectable compound to said target compound, cell or particle, said targeting group selected from the group consisting of protein, peptide and antibody:

A 2′ is a linking group or covalent bond;

B 2′ is a linking group or covalent bond;

B 2 is a water-soluble group containing at least one ionic or polar group selected from the group consisting of hydroxy, alkoxy, carboxy, nitro, cyano, amino, amido, ureido, sulfonamido, sulfinyl, sulfhydryl, silyl, S-sulfonamido, N-sulfonamido, C-carboxy, O-carboxy, C-amido, N-amido, sulfonyl, phosphono, morpholino, piperazinyl, tetrazolo, alcohol, thiol, polyethylene glycol, polyol, sulfonamide, carboxamide, hydrazide, N-hydroxycarboxamide, urea, metal chelates, carboxylate, sulfonate, phosphate, amine N-oxide, ammonium, uronic acid, carboxylic acid, sulfonic acid, amine, guanidinium, phosphoric acid, phosphonic acid, phosphatidyl choline, phosphonium, borate, sulfate, and combinations thereof; and

Z 2 is a porphyrinic macrocycle selected from the group consisting of porphyrins, chlorins, bacteriochlorins and isobacteriochlorins;

and wherein:

(i) each of said first and second detectable compounds have a different emission wavelength band, said different emission wavelength bands characterized by peaks that are separated from one another by at least 5 nanometers; or

(ii) said second compound has a lifetime at least 20 percent greater than said first compound.

11. The method of claim 10 , wherein said detecting step is carried out by flow cytometry.

12. The method of claim 10 , wherein

both of said excitation wavelength bands have a peak between 350 and 900 nanometers; and

both of said emission wavelength bands have a peak between 550 and 1000 nanometers.

13. The method of claim 10 , wherein said different emission wavelength bands are characterized by peaks that are separated from one another by at least 50 nanometers.

14. The method of claim 10 , wherein said different emission wavelength bands are characterized by peaks that are separated from one another by 10 to 50 nanometers;

and wherein each of said different emission wavelength bands have a full width at half maximum peak of not more than 30 nanometers.

15. The method of claim 1 , wherein said water soluble group is selected from the group consisting of alkoxy, carboxy, and polyethylene glycol.

16. The method of claim 10 , wherein each said water soluble group is selected from the group consisting of alkoxy, carboxy, and polyethylene glycol.