Coulter Corporation

Kenneth H Kortright

David E Hofheinz

Wallace H Coulter

Carlos M Rodriguez

Thomas R Russell

Ronald D Paul

Paulette E Smariga

Meryl A Forman

Gary P Toedter

Song Y Lee

Candie S Stoner

John D Hollinger

Constance M Hajek

R Graham Smith

Stephen F Healy, Jr

James Carey Hudson

Robert H Raynor

David M Allman

Scott M Malinconico

Carole Sullivan

A method and apparatus for accelerating at least one definitive biological reaction including increasing the accuracy of determinations made therefrom. The reaction involves selected viable biological cells which are prepared in a small sample volume and rapidly mixed with microspheres having antibody specific at least to specific ones of the cells bound thereto. The microspheres can be magnetic and the bound cells can be magnetically removed to analyze the remaining blood cell populations. The microspheres can be introduced sequentially or simultaneously.

Method and apparatus for rapid mixing of small volumes for enhancing

A method and apparatus for automatically and rapidly, retrieving, counting and/or analyzing at least one selected white blood cell population and/or subset thereof of a whole blood sample or portion thereof. A volume of a biological sample containing the white blood cells is prepared and at least one reactant specific or preferential at least to some selected biological cells is introduced thereto and rapidly mixed for a short period of time. The opacity and/or volume parameter of the cells can be modified and the mixture is then counted and analyzed in one or more steps to obtain the desired white blood cell population analysis. The biological sample can be a whole blood sample and the reactant can include or be a lyse or a monoclonal antibody bound to microspheres, which will bind to specific ones of the cells or a combination of lyse and microspheres with antibody bound thereto. The microspheres can be magnetic and the bound cells can be magnetically removed for retrieving and analyzing the remaining blood cell population.

Automated analyzer for screening cells or formed bodies for enumeration

A method and apparatus for automatically and rapidly, retrieving, counting and/or analyzing at least one selected population of cells or formed bodies, such as a white blood cell population and at least one subset thereof of a whole blood sample or portion thereof. A volume of a biological medium containing the white blood cells is prepared and at least one reactant specific or preferential at least to some selected biological cells is introduced thereto and rapidly mixed for a short period of time. The opacity and/or volume parameter of the cells can be modified and the mixture is then counted and analyzed in one or more steps to obtain the desired white blood cell population and subset analysis. The biological sample can be a whole blood sample and the reactant can include or be a lyse or a monoclonal antibody bound to microspheres, which will bind to specific ones of the cells or a combination of lyse and microspheres with antibody bound thereto. The microspheres can be magnetic and the bound cells can be magnetically removed for retrieving and analyzing the remaining blood cell population.

Method and apparatus for screening cells or formed bodies with

Method for screening cells or formed bodies for enumeration of

A method of lyophilizing mammalian cells to produce preserved human cells, hybridoma cell lines, tissue cells, and control cells for immunoassays and other hematological measurements. Prior to freezing and lyophilizing, the prepared mammalian cell pellet is suspended in a solution of trehalose in an isotonic fluid prepared at a specified optimal concentration and incubated at room temperature for a designated time period. The lyophilized cells when rehydrated, retain their optimal physiological characteristics suitable for use as an analytical control and retain said characteristics after storage at 2.degree.-8.degree. C. for a period in excess of five (5) months.

Stabilized lyophilized mammalian cells and method of making same

A hybrid cell line is provided which is capable of producing monoclonal antibodies which bind to HIV core antigen P55, the precursor protein coded for the gag gene, the core protein P24 and partial breakdown products P39 and P33. The monoclonal antibody embodying the invention does not bind the P18 core protein or any HIV envelope antigens. The cell line of the invention was developed by a unique immunization protocol in which BALB/c mice were immunized over a series of multiple infusions using a select group of immunogens and a conventional myeloma cell line for fusion with the murine splenocytes harvested. The monoclonal antibody is identified as the KC-57 antibody. This monoclonal antibody is especially useful for a solid phase immunoassay in which the monoclonal antibody is detected in its binding to HIV antigens found in a serum or plasma sample from a human patient. The cell line which produces the KC-57 monoclonal antibody has been deposited in the American Type Culture Collection, Rockville, Md. and assigned A.T.C.C. No. HB 9585.

Monoclonal antibody specific to HIV antigens

A solid-phase immunoassay is provided for determination of HIV antigens in human physiological fluid. The immunoassay is characterized by the coating of a solid substrate with a unique monoclonal antibody which recognizes a common antigenic determinant of a group of HIV core antigens and no HIV envelope antigens of HIV. The test sample preferably also is subjected to a lysing reagent prior to the incubation for uniformly dispersing antigens which may be present in the test sample.

Enzyme immunoassay for detecting HIV antigens in human sera

A solid-phase immunoassay is provided for determination of members of an immunological pair such as antigen and/or antibody of a single binding pair in human physiological fluid, wherein a single immunoassay enables simultaneous detection of antigen and/or antibody of a single binding pair in a test sample which may have circulating antigen and/or antibody. The immunoassay is characterized by the addition of an amount of antigen or 'spike' of the binding pair to the test sample prior to incubation of the test sample in the presence of a solid-phase absorbed antibody of the binding pair. The test sample preferably also is subjected to a lysing reagent prior to the incubation for uniformly dispensing antigens which may be present in the test sample.

Simultaneous enzyme immunoassay for detecting antigen and/or antibody in

A hybrid cell line capable of producing monoclonal antibodies uniquely specific to human neutrophils and eosinophils and exhibiting no specificity for lymphocytes, basophils and monocytes. Further there is noreactivity with acute leukemia cells. One of the partners in the hybridoma fusion of a mouse spleen cell developed from using human granulocytes as the immunization agent. The monoclonal antibody further is capable of being used to enumerate and isolate neutrophils in normal peripheral blood and possibly in blood of patients with acute leukemia.

Monoclonal antibody specific to a common determinant site of neutrophils

A chemical agent is provided for significantly preventing or blocking non-specific staining or binding of an antibody specific for Terminal deoxynucleotidyl Transferase (TdT) during immunofluorescent or immunoperoxidase assay procedures. These procedures include both immunofluorescent and immunohistochemical staining of samples followed by flow cytometric and/or microscopic analysis, respectively. The invention is practiced by selective use of casein introduced into the assay procedures at an appropriate interval prior to analysis using a labelled or tagged monoclonal antibody specific to a TdT epitope. The casein utilized successfully was obtained from a large variety of sources and includes the use of a non-fat milk product.

Chemical blocking agent against non-specific binding or staining of an

A murine monoclonal antibody which selectively binds to the determinant site or specific epitope of glycophorin A exposed on the membrane of the erythrocyte and does not bind to other glycophorins. The monoclonal antibody is coated on a microsphere or substrate of a suitable monodispersed variety and utilized in a separation procedure for recovery of white blood cell subsets without lysing of erythrocytes. The bound microspheres are recovered without adverse depletion of the white blood cell population of the sample. The microsphere or bead-to-cell ratio employed is most acceptable for commercial applications of the invention. The invention thus enables accurate assaying of white blood cell subsets in circulating peripheral blood without resorting to lysing of erythrocytes.

Monoclonal antibody for recovery of leukocytes in human peripheral blood

Detection of an identified human carcinoma tumor antigen in a pathological sample by means of a labelled monoclonal antibody specific to the determinant site on the antigen is enhanced and/or accelerated at an earlier development stage than heretofore achieved by removing a carbohydrate steric hindrance for monoclonal antibody availability to bind the antigen of the tumor for which it is specific. The carbohydrate steric hindrance for monoclonal binding to the antigen is identified as sialic acid. The method of the invention involves selective removal of sialic acid from the antigen's determinant site by enzymatic digestion using neuraminidase.

Method for enhancing and/or accelerating immunoassay detection of human

A murine monoclonal antibody specific for an antigenic determinant on the surface or in the cytoplasm of human carcinoma cells and tissue. A cell line is provided for producing such specific monoclonal antibodies for the detection, diagnosis, and therapeutic treatment of a plurality of human carcinomas by means of selective labelling of said monoclonal antibodies.

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Cooper City, FL, United States of America

Kenneth H Kortright