Abstract: Disclosed is a new mouse-mouse hybridoma tumor cell line A.T.C.C. No. HB8209. A monoclonal antibody produced by said cell line is specifically immunologically reactive with erythropoietin and with a polypeptide whose amino acid sequence is substantially duplicative of a sequence extent in erythropoietin. Disclosed also are procedures for isolation of erythropoietin by affinity purification and for quantitative detection of erythropoietin in fluid samples.

Abstract: Langmuir-Blodgett films having hydrophobic surfaces are chemically modified to convert the hydrophobic surfaces to hydrophilic surfaces for immobilization of active moieties having bioactive, immunoactive, thermoactive, electroactive, optoactive or redoxactive properties. Langmuir-Blodgett films having a hydrophobic surface of omega unsaturated covalent bonds are formed from an amphiphilic, bifunctional surface active material having a hydrophobic tail group bearing an omega terminus of double or triple bonded unsaturation. The amphiphilic material may be fatty acids, phospholipids or porphyrins, and is preferably omega-tricosenoylamide or omega-tricosenoic acid. Chemical modification is carried out by exposing the hydrophobic surface to a reagent such as alkaline or acidic potassium permanganate, potassium dichromate or osmium tetroxide which oxidizes the omega unsaturated covalent bonds to provide hydrophilic groups such as hydroxyl or carboxylic acid groups.

Abstract: An antibody matrix immunoassay device for determination of number and proportion of subpopulations of leukocytes is described. The device can be used to evaluate and monitor the immune status of an individual and to diagnose and monitor the progression of AIDS.

Abstract: A method for the quantitative determination of antigen (or antibody) which comprises adding a sample containing an antigen (or antibody) to a dispersion of an insoluble carrier of fine particle size with an antibody (or antigen) fixed thereto to effect an antigen-antibody reaction, measuring absorbance of the reaction mixture, A.lambda..sub.1 and A.lambda..sub.2, at two different wavelengths, .lambda..sub.1 and .lambda..sub.2, and calculating the concentration of said antigen (or antibody) in the sample from the absorbance ratio A.lambda..sub.1 /A.lambda..sub.2. This method is suitable for the measurment of the amount of an antigen or antibody in samples taken from living bodies using a simple procedure which has high sensitivity.

Abstract: The invention concerns unique immobilized immunoglobulin-binding protein materials which have a high binding capacity for immunoglobulins. Exemplified are preparations which have a high binding capacity for IgGl immunoglobulins. The preparations are made by covalently joining an immobilization support material to (a) an arginine-containing linker and (b) an immunoglobulin-binding protein material. The immunoglobulin-binding protein can be joined to the linker through an amide bond. Specifically disclosed is an immobilized protein A preparation. This immobilized protein A preparation has utility in the art of purifying monoclonal antibodies.

Abstract: A method for attachment of glycocompounds and glycoconjugates to hydrazide gels, wherein a solution of glycocompound or glycoconjugate containing a suitable oxidizing agent is brought into contact with the hydrazide gel without prior removal of the oxidizing agent. Suitably, the solution is introduced into a cartridge containing the hydrazide gel. In accordance with a first embodiment, this method requires relatively short incubations of about 30 minutes for oxidation of the glycosaccharide moiety of the glycocompound or glycoconjugate, followed by about 15 minutes for binding the oxidized glycocompound or glycoconjugate to the hydrazide gel. Pursuant to another embodiment, incubation with the oxidizing agent is carried out simultaneously with bringing the solution into contact with the gel (e.g., within the cartridge). Synthetic polymer gels with terminal hydrazide groups are particularly suitable for use in this method, as these gels are especially resistant to oxidizing agents such as sodium periodate.

Abstract: An agent or device for the determination of antigen or antibody comprising capsules of a water-insoluble, dye-permeable polymer or a liquor-pool covered by said polymer, an antien, antibody or hapten immobilized on the surface of said polymer, and a dye solution enclosed or held in said capsules or liquor-pool, and a method for determining antigen or antibody using said agent or device, which can be conveniently used for the serodiagnosis of various diseases.

Abstract: A method and test kit for detecting a first antibody using a gold particle labeled secondary antibody is described. Microspheres coated with an antigen reactive with the first antibody is reacted with the first antibody from serum or other sources. The gold-labeled antibody is reacted with the first antibody antigen complex on the microsphere and detected. Preferably the gold particles are detected using an electron microscope.

Abstract: A solid support useful for bioaffinity and ion-exchange separations and enzyme immobilization is provided. The support is based on a non-perfluorocarbon solid carrier core coated with a nonionic fluorosurfactant-coated fluorocarbon interlayer to which a ligand or a binder for the ligand is securely, but reversibly attached through a reactive perfluorocarbon anchor compound. Also provided is a solid support useful for size exclusion separations. Such support is based on a non-perfluorocarbon carrier core coated with a nonionic fluorosurfactant-coated fluorocarbon interlayer.

Abstract: A method for the detection of the presence of anti-viral or anti-bacterial antibodies in a specimen and a kit for the same. The method comprises the use of a solid phase support dipstick having attached thereto suitable antigen-binding material to which viral antigens or bacterial antigens are bound. The antibodies specific for the bound antigens bind to the antigen and a second-antibody labelled with a means for visually detecting those antibodies which have formed immunocomplexes with the bound nitrogen when in the presence of suitable reactive agents. Also, a method of using such a dipstick in this method and in the test kit is disclosed. Specifically, boving leukemia virus is tested for; a dipstick attached to a removable closure is employed; and the dipstick has two antigen binding areas, one of which is a control.

Abstract: Active agents such as proteins are covalently immobilized on substrates carrying hydroxyl groups. A silane is bound to the substrate and coupled to a heterobifunctional crosslinker at one functional group leaving a free functional group, different than the first group, to which a protein is bound while retaining high protein functionality. Preferably, the silane has a functional group which reacts with the hydroxyl group of the substrate and a thiol terminal group which reacts with a functional group of a heterobifunctional crosslinking agent which contains a succinimide group that reacts with an amino group of the active agent. Bound active agents such as proteins are useful as biosensors or reactants in a variety of applications including bioassays.

Abstract: A general method of assay for biological molecules using daylight fluorescent particles. The method described is applicable to assays involving immunological reagents, nucleic acids, hormones and neurotransmitters.

Abstract: A unitary multiple electrode sensor for detecting analytes in test samples and devices for using them. The unitary multiple electrode sensor includes a sensor support member and at least one electrode array being deposited on the sensor support member, the electrode array having sensor-activating chemical(s) attached to an embodient thereof. A test sample is applied to the electrode array to begin the test; and a sensor apparatus measures the test reaction which is occurring on the electrode array.

Abstract: Novel methods for covalent attachment of antibodies, antigens, or other molecules to solid phases using extended length heterobifunctional crosslinking reagents are disclosed. The resulting derivatized solid phases can be used in diagnostic assays.

Abstract: The present invention provides a process for the preparation of a specifically bindable protein substance bound to an insoluble carrier material, especially for use in a heterogeneous analysis process, wherein a soluable protein with a molecular weight above about 500,000, which is more hydrophobic than the specifically bindable substance, is coupled to the specifically bindable substance and then the conjugate of reaction component and protein is adsorbed on a hydrophobic solid phase.The present invention also provides a carrier material for use in solid phase immunoassays including a hydrophobic solid phase which is adsorbed on a conjugate of a hydrophobic protein with a molecular weight above about 500,000 and of a specifically bindable protein or protein containing substance.

Abstract: Homogeneous cyclophilin, a soluble binding protein, having a specific binding activity of above 50 ug cyclosporin A per mg protein and a molecular weight of about 17,600 daltons, reversibly binds immunosuppressants or antibodies thereto such as cyclosporin or anti-cyclophilin. It is isolated from the cytosol of several different mammalian tissues and can be used in various diagnostic and purifications procedures.

Abstract: An immobilized hapten reagent for use in a specific binding assay for the determination of a hapten or binding analog thereof in a liquid test sample and methods for preparing and using the immobilized hapten reagent. The immobilized hapten reagent comprises a hapten moiety covalenty linked substantially only to the external surface of a gel particle. The immobilized hapten reagent is prepared by forming a reaction mixture comprising the hapten moiety and the carrier material in a nonswelling solvent wherein the gel particle is substantially impervious to the hapten moiety and wherein an analytically insignificant amount of the hapten moiety is nonspecifically bound to the gel particle. The immobilized hapten reagent is characterized by being substantially stable in aqueous solutions and exhibits an insignificant amount of leakage of the hapten moiety into a liquid test medium.

Abstract: Fixation of a physiologically active substance on a carrier through an alkylene oxide chain allows the physioactive function of the substance to be retained to a high degree. The fixed physiologically-active substances are useful for separation and purification of materials. The carrier is preferably a thin fiber of 1.0 denier or less, and may be formed from a polymer. The alkylene oxide chain has an amino or epoxy group at one end which bonds to the carrier and a functional group at the other end which bonds to a physiologically-active substance.The carrier is preferably a thin fiber of 1.0 denier or less, and may be formed from a polymer. The alkylene oxide chain has an amino or epoxy group at one end which bonds to the carrier and a functional group at the other end which bonds to a physiologically-active substance.

Abstract: Avidin and streptavidin modified water-soluble polymers are provided, such modified polymers being exemplified by, but not limited to, water-soluble polyacrylamides being substituted by multiple substituents of avidin or streptavidin. The avidin and streptavidin modified polymers provided, may be stored, and later used to bind biotinylated antibiodies, biotinylated toxins, or biotinylated isotope-labelled proteins, thus producing homo- or heteroconjugates of known composition. A process for the preparation of certain of the avidin or streptavidin modified polymers and conjugates thereof is also provided.

Abstract: An improved assay method for detecting the presence of an antibody capable of binding with an antigen of a virus is provided. The improvement comprises using a non-human immune antibody which is reactive with an anti-human antibody as a positive control in the assay. Non-human immune IgM and a method of producing the IgM is also provided.

Abstract: Physiologically active substances are immobilized on an inorganic support, by treating the inorganic support with an aminoalkylakoxysilane having the general formula(RO).sub.3 SiCH.sub.2 CH.sub.2 CH.sub.2 NH(CH.sub.2).sub.n NH.sub.2wherein, R is an alkyl group having 1 to 4 carbon atoms, and n is an integer having a value of 5 to 12, and chemically bonding a physiologically active substance by means of an amino group to the support.

Abstract: Novel methods for covalent attachment of antibodies, antigens, or other molecules to solid phases using extended length heterobifunctional crosslinking reagents are disclosed. The resulting derivatized solid phases can be used in diagnostic assays.

Abstract: A quartz crystal microbalance assay in which the binding of analyte to a surface on or near a quartz crystal microbalance (QCM) is detected by a conjugate which comprises an enzyme capable of catalyzing the conversion of a substrate to a product capable of accummulating on or reacting with a surface of the QCM leading to a mass change and, hence, a change in resonant frequency.

Abstract: Polynucleotide sequences in a sample of biological or nonbiological material are detected by a method involving fixing of the sequences on a solid support and forming an entity between the fixed sequences and chemically-labeled polynucleotide or oligonucleotide probes having a sequence complementary to the fixed sequence for determining the identification and/or presence of the target polynucleotide sequences. The chemical label covalently or noncovalently attached to the probe comprises a signalling moiety capable of generating a soluble signal detectable by spectrophotometric assay techniques.

Abstract: An immunoassay test device is disclosed with dried reagent drops applied to a sloping side wall of a reaction wall. A method of applying them as liquid drops is also described, wherein the slope of the side wall, and the composition and volume of the liquid drop are selected to insure that the liquid drops do not flow down to the bottom of the well.

Abstract: A novel class of compounds, methods for the preparation thereof and the use thereof in chromatographic methods for binding various biologically active materials non-covalently are disclosed. The class of compounds comprises the reaction product of a polymeric gel with a pyridine base, such as 4-dimethylaminopyridine (DMAP), and a halogen-substituted pyridine, such as 3,5-dichloro-2,4,6-trifluoropyridine (DCTFP), which reaction product may in turn be optionally reacted with hydroxyl ions or specified low-molecular-weight compounds. These compounds are capable of selectively and efficiently binding proteins and other organic materials of interest non-covalently to a degree comparable or superior to the heretofore preferred natural affinity ligands, such as Protein A gels. The novel compounds find particular utility in purification and recovery of proteins such as serum albumin and immunoglobulins of various classes from crude sources, such as diluted serum samples from various species.

Abstract: The present invention provides an enzymatically-inactive, immunologically-active .beta.-galactosidase mutein, wherein, in the region between the amino acids 430 and 550, at least one amino acid of the natural sequence is changed to another amino acid and the enzymatic activity does not amount to more than 1%, referred to the native enzyme. The present invention also provides a process for the production of this mutein. Furthermore, the present invention is concerned with the use of this mutein in the immunological determination of serum proteins by the enzyme immunoassay principle.

Abstract: A unitary multiple electrode sensor for detecting analytes in blood or blood component samples and devices for using them. The unitary multiple electrode sensor includes a sensor support member and at least one electrode array being deposited on the sensor support member, the electrode array having sensor-activating chemical(s) attached to an embodiment thereof. A test sample is applied to the electrode array to begin the test; and a sensor apparatus measures the test reaction which is occurring on the electrode array.

Abstract: A substantially pure antigen found on normal and benign breast epithelial cell membranes and in breast cancer cells, fused cell hybrids which produce antibodies specific for such antigen, the monoclonal antibodies produced by such fused cell hybrids, a method for detecting the presence of breast cancer in a patient which is based on measuring the concentrations of one or more determinants of such antigen in a patient sample, and a method for either identifying those breast cancer patients whose tumors would respond to estrogen manipulation or determining prognosis based on the degree of differentiation, which method is based on measuring the concentration of an estrogen-modulated determinant of such antigen.

Abstract: A bioaffinity separation method is provided along with a solid affinity support utilized in that method. Additionally, immobilized enzyme systems are provided for use as enzyme electrode systems. The support is based on an inert perfluorocarbon polymer carrier with ligands or binders attached to its surface through a highly fluorinated isocyanate anchor group. Methods for preparing such supports and their use in capturing target molecules from samples and in analytical applications are also provided.

Abstract: A member of a bioaffinity binding pair is immobilized on a plastic surface by coating the surface with poly(ethyleneimine) derivatized with a hydrophobic group, and covalently coupling the member to the coated surface. The immobilized member may be used in immunoassays or bioaffinity separations. The derivatized poly(ethyleneimine) is preferably tosyl poly(ethyleneimine).

Abstract: Antibodies which are immobilized and covalently bound to a matrix polymer by means of a modification in a carbohydrate region of the antibodies; wherein the binding of the antibodies is effected by condensing at least one aldehyde group in an oxidized carbohydrate region and at least one epoxide function of an epoxy-group-containing matrix polymer, said condensation being conducted in the presence of a bifunctional reagent which has, on one end position of a spacer unit having at least three members, an amino group capable of condensing with the aldehyde group, and which has on the other end position a group which reacts covalently with the epoxide function.

Abstract: Immunoglobulins are purified by adsorption upon and adsorbent therefor using a buffer having a pH value of about pH 6 to pH 10 and containing at least one polycarboxylic acid in a concentration of about 0.5 M to about 0.9 M.

Abstract: Certain polyethyleneimines are used to coat solid carriers used in immunoassays to improve the sensitivity of the assay over what is obtained without the coating. Use of the invention in enzyme immunoassays yields sensitivities equal to that of radioimmunoassays. A post-coating is also used after the solid carrier has had a layer of capture molecule applied and before the assay is performed, to improve even further the sensitivity, reliability and dynamic range of the assay results.

Abstract: There is provided a novel method of testing for the presence of an analyte in a fluid suspected of containing the same. In this method, in the presence of the analyte, a substance capable of modifying certain characteristics of the substrate is bound to the substrate and the change in these qualities is measured.While the method may be modified for carrying out quantitative differential analyses, it eliminates the need for washing analyte from the substrate which is characteristic of prior art methods.

Abstract: Novel chromophor derivatives of cyclic anhydrides are provided which have the ability to react with a variety of organic substrates forming adducts which are useful in analytical techniques for the detection and measurement of biological compounds.

Abstract: An immunoassay for an analyte is disclosed in which (a) sample, (b) an immobilized specific binding partner for the analyte and (c) the specific binding partner for the analyte conjugated to a detectable marker are reacted together in a single step. Competitive interference of the reagents is avoided by using reagent (c) in a slow-release form as a glaze coated on a surface in contact with the reaction liquid.

Abstract: A bridging molecule carrying a drug, or a label such as a fluorophore, which adds across disulfide bonds of molecules, particularly proteins, and methods of manfacturing and using the bridging molecules, are disclosed. The bridging molecule is reactive with sulfhydryl groups formed by the reduction of disulfide bonds of the protein. The functional groups of the bridging molecules are typically --SH groups.

Abstract: Thiol gels are prepared from hydroxyl containing polymers by forming a 2-fluoro-1-methylpyridoxy derivative of the polymer followed by reacting the derivative with sodium dimethyldithiocarbamate and reducing to produce a sulfhydryl substituted polymer. Alternatively, the derivative is reacted with dithiothreitol to produce a sulfhydryl substituted polymer having free sulfhydryl groups spaced from the polymer. The thiol gel can be activated by means of 2,2'-dipyridyl disulfide and reacted with a free sulfhydryl containing biologically active ligand in order to provide an insolubilized ligand which can be used for various purposes including use as an immobilized biologically active material or use as a matrix in covalent chromatography.

Abstract: A porous chemically activated medium having a low affinity for peptide group-containing materials is provided comprising a porous polymeric medium having a low affinity for peptide group-containing materials covalently bound to a residue of an activating agent which is capable of reacting with an acceptor molecule.

Abstract: A bioaffinity separation method is provided along with a solid affinity support utilized in that method. Additionally, immobilized enzyme systems are provided for use as enzyme electrode systems. The support is based on an inert perfluorocarbon polymer carrier with ligands or binders attached to its surface. The ligand, binder or enzyme is preferably modified by attaching a perfluorocarbon anchor group, and the modified ligand, binder or enzyme is attached to the carrier through the anchor group. Methods for preparing such supports and their use in capturing target molecules from samples and in analytical applications are also provided.

Abstract: For the determination of a reaction partner of an immunological reaction according to the principle of immunoassay, the reaction partner to be determined is brought into contact with a marked specific receptor R.sub.1 and at least 1 unmarked receptor R.sub.2, one of the unmarked receptors R.sub.2 being bonded on a solid phase by a binding-capable substance R.sub.3. In order to determine the sample blank value, the unmarked receptor R.sub.2, which is bonded by the receptor fixed on the solid phase, is then replaced by another unmarked receptor R'.sub.2 which does not react with the reaction partner of R.sub.2 to be determined.

Abstract: A device useful for the determination of a ligand in an aqueous liquid comprises a water-insoluble substrate. Immobilized in at least one location on that substrate is a biotinylated receptor for the ligand. This receptor is in dried form and immobilized with one or more dried water-soluble acrylamide homo- or copolymer binder materials. When the immobilized receptor is contacted with an aqueous sample, the binder material is dissolved and the receptor is released for reaction with the ligand. The resulting reaction product is determined in a suitable manner. The device can be included in a kit having other components useful for ligand determination. The device and method of its use are particularly useful for the determination of human chorionic gonadotropin (hCG) as an early indicator of pregnancy.

Abstract: Macromolecular monoclonal antibody compositions are provided which are capable of selectively forming stable bonds to cells having a predetermined concentration of at least one surface antigen, such concentration being greater in such cells than in other cells in the cell population, wherein the composition comprises a substrate and a plurality of monoclonal antibodies specific to said surface antigen or antigens, which antibodies are covalently bonded to the substrate.

Abstract: There are provided magnetic and non-magnetic agarose and agar polyaldehyde beads with diameters ranging from 40 microns up to 1 cm, and processes for the synthesis of such beads. The polyaldehyde compounds e.g. polyacrolein, polymethacrolein or polyglutaraldehyde, were used as microspheres or as powders. The agarose-polyaldehyde beads are capable of covalently binding in a single step, through their aldehyde groups, compounds containing primary amino groups or thiol groups, such as proteins, antibodies, enzymes and drugs. The beads are useful for various biological applications e.g. affinity chromatography, hemoperfusion, ion exchange resins, cell labeling, diagnostic purposes and cell separation.

Abstract: Antibody coated gold sol particles and antibody coated solid phase particles dispersed in an aqueous system react immunologically as a function of the presence of an analyte in a sample to be analyzed to produce a collectible, solid phase, gold-containing composite. The composite is collected on a filter or at the bottom of a test tube by centrifugation or gravitation and the analyte in the sample is determined or detected by direct visual examination of the collected solid phase composite which has a pink or red or purplish coloration as a result of the gold contained thereby. The materials required for conducting the assay comprise coated gold particles, coated solid phase particles and a collector element such as a filter. The collected, solid phase, metal-containing composite, which may be directly visually examined to determine or detect the gold therein and thus the analyte in the sample, is stable and remains available for confirmation of test results at a later time.

Abstract: A detectable molecule of the formulaA.sup.3 --(--X--R.sup.1 --E--Det.sup.b).sub.mwhere A.sup.3 is A.sup.2 or a polymer, where A.sup.3 has at least one modifiable reactive group selected from the group consisting of amino, hydroxy, cis OH, halides, aryl, imidazoyl, carbonyl, carboxy, thiol or a residue comprising an activated carbon; --X-- is selected from the group consisting of ##STR1## --R.sup.1 -- is ##STR2## or a C.sub.1 -C.sub.10 branched or unbranched alkyl or aralkyl, which may be substituted by --OH; --Y-- is a direct bond to --E--, or --Y-- is --E--R.sup.2 -- where R.sup.2 is a C.sub.1 -C.sub.10 branched or unbranched alkyl; Z.sub.a is chlorine, bromine or iodine; E is O, NH or an acyclic divalent sulfur atom; Det.sup.b is a chemical moiety capable of being detected, preferably comprising biotin or a metal chelator of the formula: ##STR3## or the 4-hydroxy or acyloxy derivatives thereof, where R.sup.3 is C.sub.1 -C.sub.4 alkyl or CH.sub.

Abstract: A method for identifying and quantifying allergen specific IgE levels in patent serum by conjugating the IgE in the serum with allergens adhering to an insoluble support, conjugating the serum IgE with an enzyme labeled anti-IgE antibody, contacting the enzyme label with a solution of a substrate which will yield a fluorescent product in the presence of the enzyme, and measuring the level of fluorescence in the solution. Special reagents and their manufacture are also disclosed.

Abstract: A test kit is used in an agglutination immunoassay to determine a multivalent immune species, such as Streptococcus A antigen, in a biological sample. The method includes contacting an aqueous solution of the species with an agglutination indicator reagent having receptor molecules reactive with the species to form an agglutinate of the reaction product of species and receptor. These receptor molecules are bound to polymeric particles which contain tracer molecules. The resulting agglutinate is captured on a microporous membrane which has an average pore size which is at least five times greater than the average diameter of the polymeric particles. Unagglutinated residual materials are washed through the membrane using a wash solution. This solution has a pH of from about 5 to about 10 and an ionic strength of at least about 0.25. Tracer is then determined either in the agglutinate or in the residual materials.

Abstract: A polyamide dyed with a reactive dye capable of absorbing incident light of the excitation waveband of a fluorophore or light of the emission waveband of the polyamide is provided for use in assays in which the presence or quantity of an analyte is being detected by fluorescence as a result of excitation of a fluorescent material at an excitation waveband of light and in which the excitation waveband impinges upon the polyamide.