Abstract: DNA cloning shuttle vectors, including a cosmid shuttle vector, for E. coli and Streptomyces are disclosed. Specifically, disclosed shuttle vectors pAL7002 (NRRL B-18055) and pNJ1 (NRRL B-18054) contain an E. coli origin of replication, Streptomyces replication functions, and antibiotic resistance markers for both E. coli and Streptomyces. In addition, pNJ1 contains a cos sequence. Novel 2-norerythromycin antibiotics A, B, C, and D, which were produced in a strain Streptomyces erythreus 12693-240 (NRRL B-18053) transformed by pNJ1 bearing DNA from Streptomyces antibioticus, are also disclosed. The present invention also provides a method for producing novel antibiotics. This method for antibiotic production is applied to the transformation of a blocked mutant of S. erythreus with genomic DNA from S. antibioticus but may be more broadly applied to genes of antibiotic-producing strains transformed into cells which are blocked in the pathway for production of a different antibiotic.

Abstract: A diagnostic testing device is disclosed. The testing device includes a base member and a cover member which defines an opening for the reception of a liquid specimen. A distribution wheel delivers portions of the specimen to antibiotic units which are circumferentially spaced. Indicator units are in communication with the cover member. A delivery cylinder extends upwardly from each of the antibiotic units toward a respective one of the indicator units. Relative vertical movement between the delivery cylinder and the indicator unit places the cylinder and the indicator unit into an engaging relationship. A change in coloration at an indicator unit indicates a positive reaction.

Abstract: A method and kit for determining the amount of ammonia in a body fluid. The method and kit involve contacting the fluid with glutamate dehydrogenase, alpha-ketoglutarate and nicotinamide hypoxanthine dinucleotide phosphate and determining the amount of ammonia present in the fluid.

Abstract: A precipitation reagent for use in analytical systems for the determination of hydrophobic analytes in a biological test sample, particularly analytical systems employing specific binding proteins for such analytes. The precipitation reagent precipitates interfering proteins, hemoglobin, and other interfering substances from a biological test sample while, at the same, maintaining hydrophobic analytes in solution and minimizing the denaturation of specific binding proteins, such as, for example, antibodies, which may be present in an immunoassay system. The precipitation reagent comprises a zinc salt, a glycol, and a straight or branced alcohol from about 1 to 4 carbon atoms, and may optionally contain an acid. A preferred precipitation reagent comprises zinc sulfate, methanol and ethylene glycol, and is particularly useful in a fluorescent polarization immunoassay for the determination of hydrophobic analytes, especially cyclosporine.

Abstract: Disclosed is a method of producing fusion proteins wherein one part of the fusion protein is formed from the bacterial protein CKS.

Abstract: The present invention is directed to a fluorescence polarization immunoassay for determining the phenylacetylglutamine (PAG) content in body fluids, to the various components needed for preparing and carrying out such an assay, and to the methods of making these components. Specifically, tracers, immunogens and antibodies are disclosed, as well as methods for preparing them. The assay is conducted by measuring the degree of polarization of plane polarized light that has been passed through a solution continuing sample, antiserum and tracer.

Abstract: A substituted monoethylglycinexylidide or analogue is disclosed. The xylidide or analogue has the structure of FIG. 1 of the attached drawings where M is CH.sub.2 NHCH.sub.2 CH.sub.3, CH.sub.2 CH.sub.2 CH.sub.2 CH.sub.3 or CH.sub.2 OCH.sub.2 CH.sub.3, one of Y.sup.1 and Y.sup.2 is H and the other is a protein or a fluorescein moiety chemcially bonded to the glycinexylidide or analogue moiety. Also disclosed is a method for carrying out immunoassays for MEGX, which has the structure of FIG. 3 of the attached drawings. The method for carrying out immunoassays involves using the foregoing compounds as tracers and immunogens.

Abstract: This disclosure relates to a method and reagents for determining ligands in biological fluids such as serum, plasma, spinal fluid, amnionic fluid and urine. In particular, this disclosure relates to a fluorescent polarization immunoassay procedure and to a novel class of tracer compounds employed as reagents in such procedures. The procedure disclosed combines the specificity of an immunoassay with the speed and convenience of fluorescent polarization techniques to provide a means for determining the amount of a specific ligand present in a sample.

Abstract: Apparatus and method for performing a chemiluminescence assay involving the immobilization of a chemiluminescent reaction complex to a solid, porous element. The solid, porous element is preferably treated to provide an immobilizing interaction with the chemiluminescent reaction complex wherein the chemiluminescent reaction complex is thereby immobilized to the solid, porous element. The activating and reading of the chemiluminescent reaction are separately performed by evenly distributing a concentrated chemiluminescent activating solution to form a puddle on the surface of the porous element to which the chemiluminescent reaction complex is immobilized.

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 method for obtaining a substantially pure preparation of an enzyme-antibody conjugate having an enzyme component covalently coupled to a predetermined number of an antibody component where the molecular weight of the enzyme component is substantially greater than the molecular weight of the antibody component. The method involves the electrophoretic separation of the desired enzyme-antibody conjugate from an enzyme-antibody conjugate reaction mixture comprising, as separately migratable species, at least a free enzyme component and a poplulation of enzyme-antibody conjugates comprising one or more of the enzyme component convalently coupled to one or more of the antibody component. The resulting conjugate preparations are substantially pure and are therefore particularly useful as labeled reagents in immunometric assays.

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: A method for the simultaneous determination of glucose and urea in a specimen with a single reagent system. A reagent system containing a reactant for each of glucose and urea to be determined is added to a specimen. The reagent system is reacted with the specimen such that each of glucose and urea react with their respective reactant simultaneously. Each reactant is selected such that it is capable of giving an absorbance band for glucose and urea which permits their simultaneous determination. The change in absorbance or fluorescence of the resulting reaction mixture is monitored at a plurality of wavelengths which are characteristic for each of glucose and urea.

Abstract: A dispersion of a uniformly sized population of multilamellar lipid vesicles (liposomes) encapsulating an aqueous liquid is prepared by forming a dried film of lipids on the walls of a vessel, contacting the film with an aqueous liquid in the presence of spherical contact masses such as glass beads and agitating. The liposomes have mean diameters in the range of about 150 to about 3000 nanometers and the contact masses have mean diameters of about 50 to 3,000 microns. The aqueous liquid encapsulated may contain enzymes, drugs or marker substances such as colorimetric or fluorescent dyes. A member of a immunological binding pair may be associated with the surfaces of the liposomes for carrying out immunoassays. This method allows for the use of small quantities of marker and lipid, leaves no residual solvents, allows for contact with only glass surfaces and involves no transfer of liposome preparations from lipid film drying vessels to sizing apparatus.

Abstract: A method for performing a diagnostic immunoassay by solid phase separation for digoxin. To a reaction mixture of a test sample and labeled anti-digoxin antibody, which forms a complex of any digoxin present in the test sample, is added a solid phase material having an immobilized ouabain triacetate derivative compound capable of binding any excess labeled antibody. The solid phase material is chosen to rapidly settle whereby a solid and liquid phase is formed. The liquid phase can then be extracted to measure the amount of digoxin-labeled antibody present therein. Ouabain triacetate derivative compounds possess sufficient affinity for anti-digoxin antibodies, and are therefore useful in a solid phase separation based digoxin immunoassay for settling out such antibodies without contributing to undesired background interference.

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: Apparatus for measuring electrolyute concentrations in fluid samples. The apparatus includes an ion selective electrode having a plurality of ion selective detection sites. Each site has an affinity for a preselected electrolyte of interest and generates a potential having a magnitude related to the concentration of the corresponding electrolyte in the sample. A voltage to optical transducer circuit is provided to convert the voltage differentials to optical signals having intensity related to the concentration of the electrolytes in a first embodiment, a digital code related to the concentration of the electrolytes in a second embodiment, and an optical absorption or density value related to the concentration of the electrolytes in a third embodiment. The optical signals are suitable for detection by conventional optical detector apparatus of assay instruments and may be processed using conventional two point linear interpolation techniques to determine the concentrations of the preselected electrolytes.

Abstract: Optical indicator chalcogen (selenide or sulfide) compounds responsive to oxidants, e.g., hydrogen peroxide, and methods for preparing and using such indicators. Upon oxidation the resulting intermediate undergoes spontaneous elimination of the chalcogen residue to yield a signal compound which provides an optical signal such as fluorescence. Preferred fluorogenic indicator compounds are 3-chalcogen-3,4-dihydrocoumarin derivatives and 3-chalcogen-3,4-dihydro-2-quinolone derivatives. Such indicator compounds provide highly fluorescent products upon oxidation by hydrogen peroxide and are useful in analytical systems which generate hydrogen peroxide in response to the analyte under determination.

Abstract: Novel polyamino acid based coupling agents are disclosed. These reagents are useful for conjugating proteins (e.g. antibodies to enzymes) for use in diagnostic assays.

Abstract: Analytical reagent reaction vessel and method for performing sequential analytical assay procedures to determine an analyte in a liquid test sample. The reaction vessel is in the form of a closed container having a substantially horizontal axis of rotation and incorporated with one or more analytical reagents for performing a desired analytical assay procedure. The reaction vessel is designed to provide free gravitational movement of a liquid test sample disposed therein. The analytical reagents are incorporated into the reaction vessel such that they are contacted with a liquid test sample in a desired order or sequence. A liquid test sample disposed in the reaction vessel is capable of being manipulated therein by rotating the reaction vessel about the horizontal axis wherein the liquid test sample is transported by gravity.