Cyanine dyes as labeling reagents for detection of biological and other materials by luminescence methods

• US 7,008,798 B2
• Filed: 03/22/2002
• Issued: 03/07/2006
• Est. Priority Date: 04/18/1986
• Status: Expired due to Fees

• Current Assignee: Carnegie Mellon University
• Original Assignee: Carnegie Mellon University
• Inventors: Waggoner, Alan S.
• Primary Examiner(s): Ponnaluri, Padmashri
• Application Number:
  US10/103,116
• Publication Number:
  US 20020146736A1
• Time in Patent Office: 1,446 Days
• Field of Search: 435/6, 435/7.1, 436/86, 436/11, 436/501, 436/800, 436/172, 436/111, 544/212, 544/113, 544/83, 544/465, 544/469, 548/465, 548/469, 548/150, 548/156, 514/183, 514/222.2, 514/224.2
• US Class Current: 436/501
• CPC Class Codes: C07D 209/14 Radicals substituted by nitrogen atoms, not forming part of a nitro radical
• CPC Class Codes: C07D 209/60 Naphtho [b] pyrroles; Hydrogenated naphtho [b] pyrroles
• CPC Class Codes: C07D 403/12 linked by a chain containing hetero atoms as chain links
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• CPC Class Codes: C07H 5/06 Aminosugars
• CPC Class Codes: C09B 23/02 the polymethine chain containing an odd number of >CH- or >C[alkyl]- groups
• CPC Class Codes: Y10T 436/173845 Amine and quaternary ammonium

Abstract

The present invention pertains to luminescent dyes and methods for covalently attaching the dyes to a component or mixture of components so that the components may be detected and/or quantified by luminescence detection methods. The dyes are cyanine and cyanine-type dyes that contain or are derivatized to contain a reactive group. The reactive group is covalently reactive with amine, hydroxy and/or sulfhydryl groups on the component so that the dye can be covalently bound to the component. In addition, the dyes are preferably soluble in aqueous or other medium in which the component is contained. The components to be labeled can be either biological materials, such as antibodies, antigens, peptides, nucleotides, hormones, drugs, or non-biological materials, such as polymers, glass, or other surfaces. Any luminescent or light absorbing detecting step can be employed in the method of the invention.

Claims

1. A method for labeling a component of an aqueous liquid comprising:

a) adding to said liquid containing said component a luminescent dye selected from the group consisting of cyanine, merocyanine and styryl dyes containing at least one sulphonate or sulphonic acid attached to an aromatic nucleus which dye is reactive with said component;

b) reacting said dye with said component so that said dye labels said component; and

wherein said dye has a molar extinction coefficient of at least 50,000 liters per mole centimeter, a quantum yield of at least 5 percent and absorbs and emits maximally in the 400 to 900 nm range.

2. The method according to claim 1 wherein said component is selected from antibodies, proteins, peptides, enzyme substrates, hormones, lymphokines, metabolites, receptors, antigens, haptens, lectins, toxins, carbohydrates, sugars, oligosaccharides, polysaccharides, nucleotides, derivatised nucleotides, nucleic acids, deoxynucleic acids, derivatised nucleic acids, derivatised deoxynucleic acids, DNA fragments, RNA fragments, derivatised DNA fragments, derivatised RNA fragments and drugs.