Eppendorf Array Technologies S.a.

Eppendorf Ag

Eppendorf Array Technologies Sa (eat)

Eppendorf Array Technologies

Societe De Conseils De Recherches Et D'applications Scientifiques (s.c.r.a.s.)

Universitè Catholique De Louvain

Cytec Surface Specialties, S.a.

Eppendorf Array Technologies (e.a.t.)

José Remacle

Isabelle Alexandre

Nathalie Zammatteo

Dieter Husar

Heinz Koehn

Sylvain Margaine

Sandrine Hamels

Patricia Renard

Muriel Art

Francoise De Longueville

Sven De Roeck

Sophie Dufour

Laurence Lockman

Michel Tielemans

Steven Cappelle

Denis Flandre

Luc Lindekens

Lars-Erik Peters

Konrad Beyreuther

Laszlo Hevesi

Joseph Demarteau

Rémi Pampin

Carine Michiels

Martine Redon

Laurent Jeanmart

Yves Houbion

Benoit De Becker

Sven Buelow

Sophie Burteau

Pierre-Emmannuel Lobert

David Bourgeois

Luis Moreno Hagelsieb

Wilhelm Pluester

Christelle Plennevaux

Sylvian Margaine

Nathalie Zammateo

The present invention relates to a method and device for detecting and/or quantifying one or multiple target molecules present in a solution by quantifying online their binding on specific capture molecules immobilized at different locations (spots) of a surface of an optically transparent solid support without substantial detection of target molecules present in solution. The present invention allows multiple target assays to be performed in a simultaneous detection. More particularly, the invention comprises detecting in real-time the hybridization between capture DNA molecules present on a micro-array and target polynucleotides present in solution. The invention is also related to real-time PCR of multiple targets on a micro-array.

Detection and/or quantification method of target molecules on a solid support

A process for preparing a substrate having thereon sites reactive with the reactive groups of a molecular probe, the process comprising the steps of applying to the substrate surface a material comprising one or more reactive sites having an activated ethylenically unsaturated double bond of the formula I: wherein the substituents are defined in the specification and, in a further step, covalently binding the reactive group of the molecular probe to the activated ethylenically unsaturated double bond of the reactive sites having formula I.

Substrates, preparation and use

The present invention relates to a method and apparatus for monitoring a real-time quantification of multiple target molecules during their binding on capture molecules of a micro-array. The method comprises the steps of: placing, in a chamber (), a support () having fixed upon its surface a micro-array comprising at least 5 capture molecules () being immobilized in specifically localized areas () of said support; introducing said labeled target molecules solution () into the chamber; incubating said labeled target molecules under stable and controlled temperature conditions to allow the binding between said target and capture molecules; directing an excitation light () from a light source () on the surface of the micro-array; measuring the electromagnetic light emission () from the bound target molecules in response to said excitation light in presence of the solution containing the target molecules wherein the surface of emission for a localized area is comprised between about 0.1 &#x3bc;mand about 10 mmand wherein each of the at least 4 localized areas is monitored with time with at least two measurements being done per localized area (), and processing and storing the values of the different measurements and quantifying at least 4 different target molecules present in solution using at least one measurement value for each said target.

Real-time quantification of multiple targets on a micro-array

The present invention relates to a method and apparatus for monitoring on a micro-array a PCR amplification of a nucleotide molecule being present in a solution. The method includes the steps of: providing a support having fixed upon its surface a microarray having at least a capture molecule being immobilized in specifically localized areas of the support and a reaction chamber; introducing a solution containing the nucleotide molecule into the reaction chamber and reagents for nucleotide molecule amplification and labelling; submitting the solution to at least 2 thermal cycles having at least 2 and preferably 3 different temperature steps in order to obtain labelled target nucleotide molecule by PCR amplification; performing at least a measurement of the labelled target nucleotide molecule in at least one thermal cycle by incubating the labelled target nucleotide molecule under conditions allowing a specific binding between the target nucleotide molecule and its corresponding capture molecule and measuring the light emission from the bound labelled target nucleotide molecule in response to excitation light with the solution being present in the chamber and containing the labelled target nucleotide molecule. The surface of emission for a localized area is between about 0.1 &#x3bc;mand about 75 mm. The method further includes processing the data obtained in at least one thermal cycle in order to detect and/or quantify the amount of nucleotide molecule present in the solution before the amplification.

Real-time PCR of targets on a micro-array

The present invention provides a method for capacitive detection of the presence of target sample on a substrate, which comprises the steps of: binding a target sample to selective binding sites on the substrate, the target sample being directly or indirectly labeled with conductive labels, and sensing the presence of the bound conductive labels to a binding site to thereby determine the presence of the target sample. The sensing step is carried out by a capacitive detection of the presence of the conductive labels. The present invention also provides a capacitive sensor device for determining the presence of a target sample. Conductive labels are directly or indirectly couplable to the target sample. The capacitive sensor device comprises a substrate having attached thereto a binding site able to selectively bind a target sample, a capacitive sensor element, and sensing circuitry for determining the presence of a target sample bound to the binding site by application of electrical signals to a capacitive sensor element. The capacitive sensor element comprises a set of at least two electrodes with non-conductive surfaces in a region associated with the binding site.

Method and device for high sensitivity detection of the presence of DNA and other probes

A method for obtaining an activation profile of a biological sample by disposing onto a solid support in a pre-determined spatial arrangement a subset of capture molecules able to interact with one or more activated transcription factor(s) present in the biological sample, contacting the biological sample upon the solid support under conditions allowing their interaction, monitoring signals resulting from their interaction, and providing a cellular activation profile from the detected signals.

Method and kit for the determination of cellular activation profiles

Disclosed is a system and method conducting real-time PCR. Unlabeled capture molecules of a specific design are immobilized on a solid support, and contacted with amplicons produced in one or more PCR cycles. Detection of amplicons may take place during or between the PCR cycles while the solid support is in fluidic contact with the PCR solution. In an alternate embodiment detection of the amplicons takes place when the solid support is not in fluidic contact with the PCR solution. The method is suitable for the simultaneous detection and quantification of closely homologous target molecules.

Identification and quantification of a plurality of biological (micro)organisms or their components

Disclosed is a system and method conducting real-time PCR. Capture molecules of a specific design are immobilized on a solid support, and contacted with amplicons produced in one or more PCR cycles. Detection of amplicons may take place during or between the PCR cycles while the solid support is in fluidic contact with the PCR solution. In an alternate embodiment detection of the amplicons takes place when the solid support is not in fluidic contact with the PCR solution. The method is suitable for the simultaneous detection and quantification of closely homologous target molecules.

The present invention relates to a method for obtaining an activation profile of a biological sample comprising disposing onto a solid support in a pre-determined spatial arrangement a subset of capture molecules able to interact with one or more activated transcription factor(s) present in the biology cell sample, contacting the biological sample upon the solid support under conditions allowing their interaction, monitoring signals resulting from their interaction, and providing a cellular activation profile from the detected signals.

The present invention is related to a screening, detection and/or quantification method of one or more transcriptional factor(s) (1) possibly present in a biological sample, said method comprising the steps of: The present invention is also related to the kit comprising means and media for performing said method.

Method for the screening, the detection and/or the quantification of transcriptional factors

The present invention relates to a detection and quantification method of different sets of target molecules (), being possibly present simultaneously in a biological sample by the use of a microarray present upon a solid support surface () and comprising different sets (A, B) of capture molecules (), present in different locations () of the solid support surface ().

Customized micro-array construction and its use for target molecule detection

The invention relates to a method and a kit for the specific identification and/or quantification of one or several among at least 7 organisms or parts thereof, in a biological sample being possibly contaminated by at least 4 other organisms, by detecting at least one nucleotide sequence specific of each of the organisms possibly present in said biological sample, wherein said nucleotide sequence is homologous with at least 4 other nucleotide sequences. The method comprises the steps of: amplifying the nucleotide sequences specific of said organisms into target amplified nucleotide sequences using at least 2 different primer pairs, each one being capable of amplifying at least 4 of said homologous nucleotide sequences from other organisms and having an homology higher than 85% with each of the said amplified homologous nucleotide sequences to be amplified; providing an array onto which single-stranded capture nucleotide sequences are arranged at pre-determined locations, said single-stranded capture nucleotide sequences being covalently bound to an insoluble support, via a spacer which is at least 6.8 nm in length, and wherein said capture nucleotide sequences comprise a nucleotide sequence of about 10 to 50 bases which is able to specifically bind to one target amplified sequence without binding to said other amplified homologous nucleotide sequences and presenting an homology lower than 85% with the other capture nucleotide sequences of the said other amplified homologous sequences, contacting said target amplified sequences with the array in one solution under conditions allowing hybridization of the target amplified sequences to complementary capture nucleotide sequences present on the array; and detecting and quantifying signals present on specific locations on the array; wherein the intensities of the signals in specific locations allows identification of the organisms.

Method and kit for the detection and/or quantification of homologous nucleotide sequences on arrays

The present invention is related to a method for the identification and/or the quantification of a target compound obtained from a sample, preferably a biological sample, comprising the steps of putting into contact the target compound with a capture molecule in order to allow a specific binding between the target compound with a capture molecule, the capture molecule being fixed upon a surface of a solid support according to an array comprising a density of at least 20 discrete regions per cm, each of the discrete regions being fixed with one species of capture molecules, performing a reaction leading to a precipitate formed at the location of the binding, determining the possible presence of precipitate(s) in discrete region(s), and correlating the presence of the precipitate(s) at the discrete region(s) with the identification and/or a quantification of the target compound.

Method for the identification and/or the quantification of a target compound obtained from a biological sample upon chips

A method for identifying or quantifying an organism by a detecting its nucleotide sequence among at least 4 other homologous sequences comprising amplifying nucleic acids from the organism to generate target nucleotide sequences to be detected; contacting the target nucleotide sequences with single stranded capture nucleotide sequences bound by a single predetermined link to an insoluble solid support and discriminating the binding of a target nucleotide sequence specific of an organism with a signal resulting from a hybridization by complementary base pairing between the target nucleotide sequence and its corresponding capture nucleotide sequence is disclosed. The capture nucleotide sequence is bound to the insoluble solid support at a specific location on an array having a density of at least 4 different bound single stranded capture nucleotide sequences/cm. The location of the signal on the array allows identification or quantification of the organism.

Identification of biological (micro) organisms by detection of their homologous nucleotide sequences on arrays

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Malonne, Belgium

José Remacle