Beckman Coulter, Inc.

Beckman Instruments, Inc.

Robert S Matson

Jang B Rampal

Peter J Coassin

Edwin M Southern

Charles T Caskey

Raymond C Milton

Karen E Lowe

Daniel G Morrow

A microarray assay system using an array-of-arrays plate mounted on the top surface of a vacuum fixture, and an automatic handling system that handles the plate and fixture assembly. The Aplate includes a tray formed of a flexible material and a rigid frame for mounting on the fixture. The fixture has a plurality of orifices opening at the top surface and connected to a vacuum source to create a negative pressure to hold the flexible plate against the top surface of the fixture. The automatic handling systems that can be used with the plate and fixture assembly include a microarray printing machine, liquid handling robot, hybridization and incubation devices, and microarray imaging devices.

Method and apparatus for processing biomolecule arrays

Methods for preparing a substrate for the arraying and immobilizing of biomolecules or cells are described. The methods include providing a solid support and depositing by a chemical plasma-mediated polymerization process a polymeric coating on the surface. The polymeric coating comprises at least one pendant functional group capable of attaching a biomolecule or a cell. A material of the polymeric coating is preferably selected in such a way that its surface properties enhance binding between the pendant group and the biomolecule or cell. A substrate for the immobilization of biomolecules and cells is also provided. The substrate comprises a solid support made of a polymeric material. The solid support has at least one first pendant functional group suitable for attaching a first biomolecule or a first cell on its surface. The substrate further comprises a polymeric coating deposited on the surface of the substrate. The polymeric coating increases the attachment of the first biomolecule or the first cell to the first functional group.

Polymer-coated substrates for immobilization of biomolecules and cells

The present invention is directed to the production of a sample microarray for use in detecting one or more target biopolymers in the sample. The sample microarray of this invention is formed by distributing equivalent amounts of a single sample at discrete, spatially defined locations on a substrate. Each site in the microarray, thus, has the same composition of target biopolymers. The microarray is then interrogated by one or more probes specific for one or more the target biopolymers.

Analysis using a distributed sample

An assay article for detection biopolymers contained in a sample is described. The assay article includes a substrate and a biopolymer directly adsorbed on the surface of the substrate. A plurality of biopolymers may be adsorbed on the surface of the substrate to form an array. Also disclosed is a method of making the assay article. In the preferred method, an aminated polypropylene substrate is used. A biopolymer is contacted with the aminated substrate under a condition sufficient for direct adsorption of the biopolymer on the surface of the substrate. A method of detecting a target biopolymer contained in a sample is also disclosed. In this method, a substrate is contacted with either a probe or target biopolymer under a condition sufficient for a direct adsorption of either the probe or target biopolymer on the substrate to form a probe assay article or a target assay article. Then, the probe assay article is contacted with the target biopolymer, or the target assay article is contacted with the probe biopolymer under a condition that allows the formation of a probe-target complex. Finally, the complex is detected and the presence of the complex is used as a measurement for the presence or the amount of the biopolymer target contained in the sample.

Immobilization of biopolymers to aminated substrates by direct adsorption

A conductive microplate device for the detection of target biomolecules in a sample is described. The microplate comprises an assembly of a porous substrate and a conductive layer, wherein the assembly is sealed into bottom of at least some wells of the microplate. The porous substrate has a top surface and a bottom surface. The top surface comprises a plurality of covalently attached probe biomolecules. The covalently attached probe biomolecules are reactive with the target biomolecules contained in the sample. The conductive layer, which is attached to the bottom surface of the porous substrate, is adapted to receive voltage. Microplates of the present invention can be easily adapted for use with robotic workstations. Accordingly, in one embodiment, the power supply is incorporated into a robotic arm tool for fast microplate processing.

Conductive microplate

A method of attaching unmodified biopolymers, particularly, unmodified polynucleotides, directly to a solid support is provided. The method includes the steps of (a) providing unmodified biopolymers; (b) providing a solid support having at least one surface comprising pendant acyl fluoride functionalities, and (c) contacting the unmodified biopolymers with the solid support under a condition sufficient for allowing the attachment of the biopolymers to the solid support. The unmodified biopolymers may be nucleic acids, polypeptides, proteins, carbohydrates, lipids and analogues thereof. The unmodified polynucleotides may be DNA, RNA or synthesized oligonucleotides. The DNA may be single or double stranded. A device including a solid support and unmodified biopolymers attached to the solid support by reaction with the pendant acyl fluoride functionalities of the solid support is also provided. The methods and devices of the present invention may be used in performing hybridization assays and immunoassays.

Immobilization of unmodified biopolymers to acyl fluoride activated substrates

The present invention relates to a process for detecting the presence of at least one specific nucleic acid sequence in a sample containing a nucleic acid or a mixture of nucleic acids by amplifying the nucleic acid using polymerase chain reaction, cleaving the amplified products with uracil DNA glycosylase to obtain short DNA segments and detecting the DNA fragments by using reverse blot hybridization.

Use of uracil-DNA glycosylase in genetic analysis

The present invention relates to a method of immobilizing oligonucleotides and other biomolecules using a carboxylated polyvinylidene fluoride support membrane for the construction of geosensors and other array-based systems. The support membrane may be used in either covalent or non-covalent binding states.

Carboxylated polyvinylidene fluoride solid supports for the

A solid support based hybridization assay is provided which allows for the systematic and reproducible analysis of repeat and tandem repeat oligonucleotide sequences of DNA and RNA by hybridization to a reverse dot blot array comprising strings of such repeats complementary to those found in particular nucleic acid targets (e.g., analyte PCR product). An addressable library (i.e., an indexed set) of complementary repeats is synthesized on a suitable support. Preferably, the support comprises a low fluorescent background support, thereby facilitating the use of non-radioisotopic modes of detection (such as fluorescence or chemiluminescence); particularly suitable in this regard is an aminated polypropylene support or similar material. Preferred arrays permit screening of DNA and RNA samples for complete sets of particular types of nucleotide repeat sequences (e.g., all nucleotide doublet or triplet repeats).

Oligonucleotide repeat arrays

Disclosed herein are surface activated, organic polymers useful for biopolymer synthesis. Most preferably, aminated polypropylene is used for the synthesis of oligonucleotides thereto, and these devices are most preferably utilized for genetic analysis of patient samples.

Surface activated organic polymers useful for location - specific

Disclosed herein are surface activated, organic polymers useful for biopolymer synthesis. Most preferably, aminated biaxially oriented polypropylene is used for the synthesis of oligonucleotides thereto, and these devices are most preferably utilized for genetic analysis of patient samples.

Biopolymer synthesis using surface activated biaxially oriented

An automated method and apparatus for performing biopolymer synthesis on a two-dimensional support surface whereby a two-dimensional matrix or array of biopolymers are obtained on the surface. An applicator in the form of a thick plate having in a surface at least one cavity in the form of an open chamber or channel is used for applying reagents to the surface of a solid phase support material. The applicator is positioned over the solid support material with the surface having the cavity sealed against the material surface. For each cavity, reagents for synthesis are introduced sequentially into one end of the cavity and collected from the other end of the cavity. A one-dimensional array of biopolymers are thus formed on the support material, where each element of the array contains a population of biopolymers having identical sequence. Further synthesis of additional biopolymers is performed by linearly moving or rotating the applicator relative to the support material and carrying out synthesis procedure so that another one-dimensional matrix or array of biopolymers are formed at an offset position to the previous array. At each overlapping region of the two arrays, a cell of biopolymers are formed which each comprises a strand from the first array and a strand from the second array. The result is a two-dimensional array of biopolymers of different sequences at each discrete cell. The arrays of discrete cell containing different biopolymers may be used to conduct hybridization reaction analysis of a biological sample.

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Orange, CA, United States of America

Robert S Matson