Dako Denmark A/s

Agilent Technologies, Inc.

Other

Isis Pharmaceuticals, Inc.

Jesper Lohse

Kenneth Heesche Petersen

Lars Winther

Mike Paw Maischnack Hansen

Susanne Gabs

Galina Skladtchikova

Muthiah Manoharan

John S Kiely

Henrik Pedersen

Michael Egholm

Peter Eigil Nielsen

Kelly G Sprankle

Ole Buchardt

Rohit Jain

Joachim Schmid

Liselotte Brix

Michael C Griffith

Jørgen Schøller

Tina Brink Jakobsen

Kirsten Damgaard Hoff

Galina Skladchikova

Jeffery Caron

Soren Holst Sonnechsen

Thomas Briscoe

Hans Christian Pedersen

Kivin Jacobsen

Katja Brunstedt

Lars Christian Jacobsen

Rikke Malene Krusenst Jerna-Hafstrom

Kristian Jensen

Jeff Caron

Rikke Malene JØRGENSEN

Tina Jakobsen

Rikke Malene Krusenstjerna-Hafstrom

Chromogenic conjugates for color-based detection of targets are described. The conjugates comprise a chromogenic moiety such as a rhodamine, rhodol or fluorescein. The chromogenic moiety is linked to a peroxidase substrate. The chromogenic conjugates can be used in immunohistochemical analysis and in situ hybridization. The conjugates can be used to detect 1, 2, 3 or more targets in a sample by color.

Chromogenic peroxidase substrates

The present invention relates to methods of visualizing targets in histological samples, e.g. biopsy samples, wherein the methods comprise staining of the sample with (i) one or more target specific immunochemical stains, and (ii) a histological stain for specific tissue components e.g. iron, mucins glycogen, amyloid, nucleic acids, etc., e.g. hematoxilyn and/or eosin stains or the like, that is used to enhance contrast in the microscopic image of a tissue sample, highlight morphologic structures in the sample for viewing, define and examine tissues, cell populations, or organelles within individual cells. Methods may further comprise evaluation of expression of one or more targets in the sample. The disclosed methods are useful for medical diagnostics.

Combined histological stain

The present invention lies in the field of visualization and quantification of immobilized targets in samples using immunochemical means. In particular, the invention relates to a method and reagents for detection, visualization and quantification of a molecular target in immunostained histological samples using particular compositions of the target specific binding agent. Methods and compositions of the invention are suitable for any assay that uses a target visualization system in histological samples based on detection of the target by the target specific binding agent. The methods and compositions are useful for evaluation of targets that are biomarkers of diseases in medical diagnostic.

Method for evaluation of target in histological sample

Chromogenic conjugates for color-based detection of targets are described. The conjugates comprise a chromogenic moiety such as rhodamine, rhodol or fluorescein. The chromogenic moiety is linked to a peroxidase substrate. The chromogenic conjugates can be used in immunohistochemical analysis and in situ hybridization. The conjugates can be used to detect 1, 2, 3 or more targets in a sample by color.

The present invention lies in the field of visualization and quantification of immobilized targets in samples using immunochemical means. The methods of the invention utilize an immunostaining system allowing visualizing single target units in samples as distinct dots. In particular, the invention relates to methods and reagents for visualization and quantification of molecular targets immunostained in histological samples and use of said method and reagents in medical diagnostic. However, the visualization and quantification methods of the invention are applicable to a variety of targets in different samples and allow precise quantifying both relative and absolute amounts thereof.

Quantification of single target molecules in histological samples

The present invention relates to a method of biological labeling that occurs via a free radical chain reaction. The labeling occurs due to deposition of a detectable reporter molecule from a media comprising a substance comprising at least two moieties of a peroxidase enzyme substrate (termed herein 'cross-linker') in a target site comprising peroxidase activity and a biological marker. The labeling reaction described herein may generally be used to detect targets in a host of experimental schemes for detecting and visualizing a biological or chemical target, including immunohistochemistry (IHC), in situ hybridization (ISH), antibody-based staining methods such as ELISA, Southern, Northern, and Western blotting, and others.

Rapid and sensitive method for detection of biological targets

The invention provides compositions and methods for the detection of targets in a sample; in particular, an in situ hybridization (ISH) sample. Probes and detectable labels may be provided in multiple layers in order to increase the flexibility of a detection system, and to allow for amplification to enhance the signal from a target. The layers may be created by incorporating probes and detectable labels into larger molecular units that interact through nucleic acids base-pairing, including peptide-nucleic acid (PNA) base-pairing. Optional non-natural bases allow for degenerate base pairing schemes. The compositions and methods are also compatible with immunohistochemistry (IHC), immunocytochemistry (ICC), flow cytometry, enzyme immuno-assays (EIA), enzyme linked immuno-assays (ELISA), blotting methods (e.g. Western, Southern, and Northern), labeling inside electrophoresis systems or on surfaces or arrays, and precipitation, among other general detection assay formats. The invention is also compatible with many different types of targets, probes, and detectable labels.

In situ hybridization detection method

The present invention relates to a soluble negative control MHC multimer comprising a nonsense peptide that binds the MHC protein efficiently, but that does not support binding of the resultant MHC-peptide complex to the desired T Cell Receptor. The nonsense peptide is designed to i) have a length enabling binding to the MHC allele in question, ii) have appropriate amino acids at relevant anchor positions which anchor the nonsense peptide to the peptide-binding groove of the MHC, iii) have amino acids outside the anchor positions that do not support binding to a T Cell Receptor, and iv) have an amino acid sequence that is different from the linear sequence of any naturally occurring peptide.

MHC multimers, methods for their generation, labeling and use

Disclosed are methods and systems for analyzing images of specimens processed by a programmable quantitative assay or more specifically a robust programmable quantitative dot assay, PDQA, that enable specimens to be imaged and assessed across a wide variety of conditions and applications. Specific embodiments directed to immunohistochemical applications provide more quantitative methods of imaging and assessing biological samples including tissue samples.

Methods and systems for analyzing images of specimens processed by a programmable quantitative assay

The present invention relates to methods and compounds for enzyme-mediated amplification of target-associated signals for visualization of biological or chemical targets in samples, wherein the targets are immobilized, in particular the invention relates to the oxidoreductase-mediated signal amplification for visualization of targets in samples comprising cells. The methods of the invention are particular useful for qualitative and quantitative evaluation of biological markers relating to medical diagnostic and therapy.

Method for enzyme-mediated signal amplification

The present invention relates to compounds which are conjugates of two or more moieties capable of serving as substrates of an enzyme with peroxidase activity and one or more labels. The conjugate molecules of the invention are useful for detection of molecular targets, e.g. biological or chemical molecules, molecular structures, etc, in samples using a host of experimental schemes for detecting and visualizing such targets, e.g. immunohistochemistry (IHC), in situ hybridization (ISH), ELISA, Southern, Northern, and Western blotting, etc. The invention also relates to compositions comprising the conjugate molecules.

Conjugate molecules

The present invention concerns monomeric or polymeric linker molecules useful in biological and chemical applications, their synthesis, and the synthesis and use of derivatives of the linkers conjugated to a variety of detectable labels and other substances. The linkers may be used, for example, in conjunction with fluorescent labels, nucleic acid or nucleic acid analog probes, and solid phase systems, and to enhance the solubility of the conjugated molecules.

Monomeric and polymeric linkers useful for conjugating biological molecules and other substances

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Herlev, Denmark

Jesper Lohse