Coulter Corporation

Coulter International Corp.

Alexander Burshteyn

Olavi Siiman

Ravinder K Gupta

John A Maples

Frank J Lucas

Robert H Raynor

Nazle Kuylen

Cecilia Smith

Julie Wilkinson

John W Joubran

Patricia Roth

Ravindra Mylvaganam

James Keller Whitesell

Michael L Bell

Carlos M Rodriguez

Jack D McNeal

Sandra Socarras

Carlos Aparicio

Kristie M Gordon

Meryl Forman

Paul W Price

Maria Elena Insausti

Marilyn Cayer

Orlando Concepcion

A method for utilizing a filtration device for removing interferants from a test sample containing a mixture of a composition of interest and interferants in an automated apparatus is disclosed. The filtration device includes a microporous hollow fiber membrane having a plurality of pores sized to retain the composition of interest while allowing smaller diameter interferants to pass through the membrane.

Apparatus and method for sample purification and analysis

A method for utilizing a filtration device for removing interferants from a sample containing cells in an automated apparatus is disclosed. The filtration device includes a microporous hollow fiber membrane having a plurality of pores sized to retain cells while allowing smaller diameter interferants to pass through the membrane. The apparatus also includes a means for of moving the sample from a sample container to and from the filtration device. The disclosed method utilizes a vacuum source to aspirate the sample into a lumen of the hollow fiber membrane so that the sample is retained in the lumen space until expelled into an analysis container or transported to an analyzer.

Apparatus for biological sample preparation and analysis

Ligand-aminodextran-(phycobiliprotein or tandem dye) conjugates useful for detection of a desired target biological material by providing an enhanced fluorescent signal are described. Also described is a method for a single-measurement quantification of multiple populations of cells based upon the labeling of different pairs of cell populations, each pair containing mutually exclusive cell receptors which are expressed at substantially similar receptor densities with labeled ligands for each receptor. One cell population is labeled with a ligand capable of binding to a first cell surface receptor which ligand is directly conjugated to a fluorescent phycobiliprotein or tandem dye; and a second cell population is labeled with a ligand capable of binding to a second cell surface receptor, which ligand is cross-linked to an aminodextran to a fluorescent phycobiliprotein or tandem dye.

Ligand-aminodextran-marker conjugates and uses thereof

A stable protein-coated nickel particle useful in biological assays contains a nickel particle having removed from the surface thereof nickel oxide; a linker attached to said nickel particle, the linker having a free amino group; and a protein attached to said linker by covalently bonding to the free amino group. Methods of producing and using these oxide-free nickel-protein conjugates are disclosed.

Stable protein-nickel particles and methods of production and use thereof

A method for a single-measurement quantification of multiple populations of white blood cells (WBC) is based upon the labeling of different pairs of cell populations, each pair containing mutually exclusive cell receptors which are expressed at substantially similar receptor densities with labeled ligands for each receptor. One cell population is labeled with a ligand capable of binding to a first cell surface receptor which ligand is directly conjugated to a fluorescent phycobiliprotein or tandem dye; and a second cell population is labeled with a ligand capable of binding to a second cell surface receptor, which ligand is cross-linked by an aminodextran to a fluorescent phycobiliprotein or tandem dye. The phycobiliproteins upon laser excitation produce a different detectable fluorescence intensity for each cell population. Use of such pairs of conjugates enable two populations of cells with similar receptor densities to be distinguished with the use of a single color marker. Further use of additional ligands to other cell surface receptors and additional phycobiliprotein or tandem dye, or other markers in the same manner, enables the simultaneous quantification of up to 45 different cell populations with one laser line and four fluorescent colors.

Simultaneous analyses of white blood cell subsets using multi-color,

A stable colloidal particle comprises a colloidal-sized core substrate having amine-reactive functional groups thereon with an aminodextran coating over its peripheral surface and a layer of colloidal-sized metallic solid overlaying the aminodextran coating. A linker comprising aminotrithiolate attached to the metallic solid has a free amino group to which a protein is attached by covalent bonding to the free amino group. Such novel particles are useful in flow cytometry, particularly useful in the simultaneous analyses of subpopulations of leukocytes. Methods of preparation of such particles, as well as methods of use are provided. These conjugates enable the simultaneous analyses of at least two different mutually exclusive subsets of white blood cells, based on the binding affinity of the conjugated protein.

Protein-colloidal metal-aminodextran coated particle and methods of

The present application discloses dextran crosslinked, antibody-phycobiliprotein conjugates containing up to twenty phycobiliprotein per dextran molecule which conjugates are able to achieve amplification of fluorescence intensity over direct antibody-phycobiliprotein labeled cells of two fold or greater.

Antibody-aminodextran-phycobiliprotein conjugates

This invention relates generally to using fluorescent markers (labelled antibodies) and flow cytometry to enumerate the average number of receptors (antigen) on formed bodies (cells) in whole blood, and to evaluate the specific binding constant of the marker for the particular receptors. Mean channel fluorescence intensities of equilibrated marker-cell suspension mixtures, total concentrations of marker, and targeted cell counts obtained by established procedures are used to complete the analyses. The invention further describes a competitive binding assay between marker and unlabelled antibody for receptors shed from the surface of formed bodies into solution and receptors on the formed bodies to analyze for receptors in solution. Moreover, competitive binding assays between marker and unlabelled antibody for receptors on formed bodies in whole blood are described for determination of relative, specific, and average binding constants of unlabelled/labelled antibody for targeted receptors.

Methods of enumerating receptor molecules for specific binding partners

Gelatin and aminodextran coated polymer core particles useful in immunoassays and methods of making the same are disclosed. The preparation of aminodextrans having varying amounts of amine groups is also described, as is a method of crosslinking gelatin and aminodextran without the use of a stabilizer.

Polymeric particles having a biodegradable gelatin or aminodextran

The invention relates generally to colloidal particle having a core material and a gelatin/aminodextran coating with pendent functional groups attached thereto. Biological substances or molecules, especially monoclonal antibodies, may be attached to said particles. The monoclonal antibody containing particles are useful in a variety of positive and negative biological assays.

Biodegradable gelatin-aminodextran particle coatings of and processes

The invention describes stable colloidal polymeric particles of 0.2-5.0 microns size which are coated with a first aminodextran layer and a second metallic solid layer, The metal is coated by reduction of a metallic salt or complex by the aminodextran. The metal coated particles, which preferably are gold- or silver-coated polymeric microspheres, produce side scatter and forward shifts in flow cytometry applications that are in agreement with the theoretical shifts predicted for solid gold or silver spheres of similar size in flow cytometry applications,

Immobilized metal colloids on dispersed polymer microspheres

The invention related generally to the preparation of colloidal metal(O) particles having a crosslinked aminodextran coating with pendent amine groups attached thereto. The aminodextran acts as both a reductant for reducing metal ions to metal(O) particles and as the protective agent which coats the metal(O) particles thus formed. After stabilizing the aminodextran coating by use of a crosslinking agent, the coated particles can be used to covalently bind proteins. The resulting protein containing colloidal particles can be used as markers in optical and electron microscopy, in immunological and biological assays, and possibly as therapeutic agents.

Formation of colloidal metal dispersions using aminodextrans as

A method is described for the preparation of uniform colloidal particles of ferrites, containing manganese(II), at a relatively low temperature in the presence of an aminodextran solution which acts as a support vehicle for the nucleation and growth of colloidal particles of metal oxide and for dispersion as separate single particles.

Growing community of inventors

Hialeah, FL, United States of America

Alexander Burshteyn