Chromagen, Inc.

La Jolla Pharmaceutical Company

Other

Aries Associates, Inc.

La Jolla Pharmaceutial Company

Michael J Conrad

Stephen M Coutts

David S Jones

Douglas Alan Livingston

John P Hachmann

Gene Gould

Yifeng Xiong

Ram S Bhatt

Azzouz Bencheikh

Liyan He

The present invention provides novel fluorescent compounds and covalent attachment chemistries which facilitate the use of these compounds as labels for ultrasensitive and quantitative fluorescent detection of low levels of biomolecules. In a preferred embodiment, the fluorescent labels of this invention are novel derivatives of the hydroxy-pyrene trisulphonic and disulphonic acids which may be used in any assay in which radioisotopes, colored dyes or other fluorescent molecules are currently used. Thus, for example, any assay using labeled antibodies, proteins, oligonucleotides or lipids, including fluorescent cell sorting, fluorescence microscopy (including dark-field microscopy), fluorescence polarization assays, ligand, receptor binding assays, receptor activation assays and diagnostic assays can benefit from use of the compounds disclosed herein.

Green and orange fluorescent labels and their uses

A fluorescence spectrophotometer having an excitation double monochromator, a coaxial excitation/emission light transfer module, and an emission double monochromator. Each monochromator includes a pair of holographic concave gratings mounted to precisely select a desired band of wavelengths from incoming broadband light without using other optical elements, such as mirrors. Selected excitation light is directed into a sample well by a light transfer module that includes a coaxial excitation mirror positioned to direct excitation light directly to the bottom of a well of a multi-well plate. Fluorescence emission light that exits the well opening is collected by a relatively large coaxial emission mirror. The collected emission light is wavelength selected by the emission double monochromator. Selected emission light is detected by a photodetector module.

Scanning spectrophotometer for high throughput fluorescence detection

A fluorescence spectrophotometer system may be implemented in scanning fluorescence polarization detection applications. A wavelength and area scanning fluorescence spectrophotometer system may include a light source, an excitation double monochromator, an excitation/emission light transfer module, an emission double monochromator, a high speed timer-counter circuit board, a precision positioning apparatus for positioning a sample relative to the focal plane of the excitation light, and polarizing filters at the excitation side and the emission side. The system may be operative to analyze more than one fluorescent compound in the sample; additionally or alternatively, the system enables analysis of samples from selected ones of a plurality of samples.

Scanning Spectrophotometer for high throughput fluorescence detection and fluorescence polarization

The subject invention provides compounds useful as fluorogenic substrates for the hydrolytic enzymes. Upon hydrolysis of the hydrolyzable group, a halo-pyrene substituted molecule is developed which is highly fluorescent, water soluble and exhibits several desirable characteristics, including a large Stokes' shift.

Fluorogenic substrates

Scanning spectrophotometer for high throughput fluroescence detection

The subject invention provides compounds useful as fluorogenic substrates. Upon hydrolysis of a hydrolyzable group, a halo-pyrene substituted molecule is developed which is highly fluorescent, water soluble and exhibits several desirable characteristics, including a large Stokers' shift.

Fluorogenic substrates and their use

Structural analogs of the six non-fluorescentN-nucleosides commonly found in RNA and DNA, which are inherently fluorescent under physiological conditions, are identified and methods for their preparation provided. Such analogs may be incorporated into DNA and/or RNA oligonucleotides via either enzymatic or chemical synthesis to produce fluorescent oligonucleotides having prescribed sequences. Such analogous sequences may be identical to, or the analogous complement of, template or target DNA or RNA sequences to which the fluorescent oligonucleotides can be hybridized. Methods of preparing either RNA or DNA oligonucleotide probes of the invention, intermediates used in such methods, and methods of using the probes of the invention in oligonucleotide amplification, detection, identification, and/or hybridization assays are also provided.

Fluorescent N-nucleosides and fluorescent structural analogs of N-nucleosides

Phosphoramidites of the formula ##STR1## where R is a base-labile protecting group, R.sup.1 and R.sup.2 are individually alkyl of 1 to 6 carbon atoms, cycloalkyl of 3 to 8 carbon atoms, or aryl of 6 to 20 carbon atoms or are joined together to form with the nitrogen atom a cyclic structure of 4-7 carbon atoms and 0 to 1 annular chalcogen atoms of atomic number 8 to 16, G is a hydrocarbylene group of 1 to 20 carbon atoms and Z is a hydroxy-protected vicinal diol group bound to G by one of the vicinal diol carbon atoms or a disulfide group and bound to G by one of the sulfur atoms of the disulfide group, with the proviso that G is of at least 4 carbon atoms when Z is said disulfide group are used in conventional automated oligonucleotide synthesis to introduce a functional aldehyde or thiol group on the 5' end of the oligonucleotide to thereby provide a reactive site on the oligonucleotide that may be used to conjugate the oligonucleotide to molecules that contain a free amino group or an electrophilic center reactive with a thiol group.

Modified phosphorous intermediates for providing functional groups on

Structural analogs of the six non-fluorescent N-nucleosides commonly found in RNA and DNA, which are inherently fluorescent under physiological conditions, are identified and methods for their preparation provided. Such analogs may be incorporated into DNA and/or RNA oligonucleotides via either enzymatic or chemical synthesis to produce fluorescent oligonucleotides having prescribed sequences. Such analogous sequences may be identical to, or the analogous complement of, template or target DNA or RNA sequences to which the fluorescent oligonucleotides can be hybridized. Methods of preparing either RNA or DNA oligonucleotide probes of the invention, intermediates used in such methods, and methods of using the probes of the invention in oligonucleotide amplification, detection, identification, and/or hybridization assays are also provided.

Applications of fluorescent N-nucleosides and fluorescent structural

Fluorescent universal nucleic acid end label

Structural analogs of the six non-fluorescent N-nucleosides commonly found in RNA and DNA, which are inherently fluorescent under physiological conditions, are identified and methods for their preparation provided. Such analogs may be incorporated into DNA and/or RNA oligonucleotides via either enzymatic or chemical synthesis to produce fluorescent oligonucleotides having prescribed sequences. Such analogous sequences may be identical to, or the analogous complement of, template or target DNA or RNA sequences to which the fluorescent oligonucleotides can be hybridized. Methods of preparing either RNA or DNA oligonucleotide probes of the invention, intermediates used in such methods, and methods of using the probes of the invention in oligonucleotide amplication, detection, identification, and/or hybridization assays are also provided.

Growing community of inventors

San Diego, CA, United States of America

Michael J Conrad