Abstract: Phosvitin or a modified phosvitin immobilised and coupled to a suitable matrix may be used for the separation and purification of proteins or polypeptides and in the removal of metal ions from biological material. If desired the phosvitin or modified phosvitin may be in the form of a metal chelate complex.

Abstract: Disclosed is an apparatus designed to be used for enriching specific cell types from cell mixtures. The apparatus includes a centrifugable device that includes a constriction defining a lower region and a defined cell separation medium. The constriction prevents mixing between the upper and lower portions of the device. Also disclosed are methods that use precisely defined cell separation media to isolate specific cells from cell mixtures, including CD34.sup.+ hematopoietic progenitor cells from blood or bone marrow, nucleated fetal cells from maternal blood, specific tumor cells, dendritic cells, natural killer cells, and natural suppressor cells from various body fluids, and for enrichment or depletion of T cell lymphocytes. Also disclosed is a density adjusted cell separation technique used to augment the above apparatus and enrichment methods. The apparatus and enrichment methods are useful in various diagnostic and therapeutic regimens.

Abstract: Hybridization method for discriminating between complementarity and partial complementarity of DNA base sequences. Kinetic covalent entrapment of PCR-amplified target DNA is achieved through the use of crosslinkable probes. A crosslinking site is introduced into the target DNA via the PCR amplification process. Problems with renaturation of target DNA in the PCR process and in the hybridization reaction are minimized.

Abstract: A novel homogeneous human cytokine, Natural Killer Stimulatory Factor, having the ability to induce the production of gamma interferon in vitro in human peripheral blood lymphocytes, and a pharmaceutical preparation containing it.

Abstract: The present invention relates to methods of enriching breast tumor cells from a patient's body fluids. In particular, it relates to the use of a cell-trap centrifugation tube containing a specific density gradient solution adjusted to a specific density to enrich for breast tumor cells from a cell mixture. The tube allows the desired cell population to be collected by decantation after centrifugation to minimize cell loss and maximize efficiency. In addition, the method can be further simplified by density-adjusted cell sorting which uses cell type-specific binding agents such as antibodies and lectins linked to carrier particles to impart a different density to the non-tumor or tumor cell populations allowing the breast tumor cells to be separated from the non-tumor cells in a more convenient manner.

Abstract: The present invention relates to methods of enriching fetal cells from maternal body fluids. In particular, it relates to the use of a cell-trap centrifugation tube containing a gradient solution adjusted to a specific density to enrich for fetal nucleated red blood cells from maternal blood. The tube allows the desired cell population to be collected by decantation after centrifugation to minimize cell loss and maximize efficiency. In addition, the method can be further simplified by density-adjusted cell sorting which uses cell type-specific binding agents such as antibodies and lectins linked to carrier particles to impart a different density to undesired cell populations allowing the fetal cells to be separated during centrifugation in a more convenient manner. The rapid fetal cell enrichment method described herein has a wide range of applications, including but not limited to, gender determination and prenatal diagnosis of genetic diseases without the use of invasive procedures.

Abstract: A process for purifying an antibody is provided. In this process, a mixture containing the antibody and contaminant is subjected to low pH hydrophobic interaction chromatography (LPHIC) optionally at low salt concentration. The antibody is eluted from the column in the fraction which does not bind thereto. This process can be preceded and followed by other purification steps.

Abstract: The disclosure describes highly specific tryptase polyclonal antibodies, and a method to purify the antibodies. Specifically, the invention relates to polyclonal antibodies which have the capacity to capture tryptase out of solution, a process to generate the antibodies, and an enzyme-linked immunosorbent assay (ELISA) for human tryptase which utilizes the antibodies.

Abstract: A method and a kit for separating double-stranded nucleic acid molecules from a mixture containing both single-stranded and double-stranded nucleic acid molecules. The method is particularly suitable for separating hybridized from unhybridized probe nucleic acid.

Abstract: Nonporous polymer beads having an average diameter of about 1-100 microns are suitable for chromatographic separation of mixtures of nucleic acids when the polymer beads are alkylated with alkyl chains having at least three carbon atoms. The polymer beads provide efficient separation of nucleic acids using ion-pair reverse phase chromatography.

Abstract: A economically viable method for producing sugars using concentrated acid hydrolysis of biomass containing cellulose and hemicellulose is disclosed. The cellulose and hemicellulose in the biomass is first decrystallized and then hydrolyzed to produce a hydrolysate containing both sugars and acid. Silica present in the biomass can then be removed for further processing. The remaining solids are then subjected to a second decrystallization and hydrolyzation to optimize the sugar yields. An improved method for separating the sugars from the acid in the hydrolysate is also disclosed. The resulting sugar stream can then be fermented, using an improved method which allows both hexose and pentose sugars to be fermented simultaneously.

Abstract: An integrated cell culture-protein purification system has been developed for the continuous, automated production of pure cell culture protein products. The instrument comprises a bioreactor subunit having a hollow fiber bioreactor to culture and maintain cells which secrete the desired product into the cell culture medium. The culture medium is circulated through a purification cartridge which adsorbs the desired product. The system is capable of continuously removing the product from the culture medium by immunoaffinity adsorption and pure product is then recovered. The product can also be recovered automatically in a discontinuous operation from the spent culture medium by either affinity, ion exchange, or hydrophobic purification techniques. The integrated system allows continuous production of high-quality, pure proteins from cell culture.

Abstract: The present invention provides an affinity separation method involving dynamic filtration.

Abstract: A biologically active, stable protein with a molecular weight of 20,000 to 30,000 and an isoelectric point of 8.0-10.0 is produced from the culture fluid of Streptococcus pyogenes and purified. The purified biologically active protein produces lymphocyte proliferation, provides protection against bacterial and viral infection and restricts minor metastasis.

Abstract: A method and kit for extracting fungus from a plant. An extraction solution is combined with tissue from a plant and the combination is agitated, preferably by shaking, to extract a detectable amount of the fungus from the plant. The extraction solution is a solution containing a dilute acid and a detergent. The extract can then be neutralized without degradation of the fungus and subjected to analysis by immunoassay to detect the presence of pathogenic fungi. The method is particularly useful for the detection of Pyricularia oryzae on rice plants.

Abstract: The amount of an analyte in a sample derived from a living sample is measured by reacting the analyte with an excess of a substance having affinity for the analyte, followed by separation of complex by high pressure liquid chromatography and measurement using a linear calibration curve representing the peak area values associated with known concentrations of analyte.

Abstract: Improvements in the existing procedures and materials for conduct of high gradient magnetic separation (HGMS) are disclosed. Superior superparamagnetic particles, optionally coated with a polysaccharide or other, usually organic, materials can be prepared in uniform compositions with homogeneous magnetizations. The coating can conveniently be conjugated to a specific binding moiety complementary to a biological material whose purification or separation is desired. In addition, plastic coated matrices which form superior magnetic gradient-intensifying supports are disclosed, along with improved methods and apparatus to conduct HGMS.

Abstract: Applicants describe methods for purifying human ApoE or analog thereof from recombinant bacterial cells with minimal protein aggregation and degradation during the purification process. The invention involves the addition of neutralized fatty acids to the medium during cell disruption and the use of a non-ionic detergent during the purification process. Additionally applicants describe a method for increasing the production of ApoE or analog thereof in a bacterial host by adding to the culture medium neutralized fatty acids, fatty acid precursors, triglycerides, triglyceride precursors or acetate. Pharmaceutical and diagnostic uses of the ApoE analog are described.

Abstract: A method for increasing the quantitation of microorganisms in cell-containing or cell-free blood samples, included those intended for blood product transfusion purposes, is provided which employs a sustained level of sodium polyanethol sulfonate throughout microbial replication on solid media. In another embodiment, purified saponin is combined with sodium polyanethol sulfonate to provide increased quantitation of microorganisms.

Abstract: A device for collecting a specimen, including (a) a test vessel for containing a solution containing an immunocomplex, which is a complex as a result of an antigen-antibody reaction between a specimen and a magnetic-labeled antibody containing a magnetic micro-particle and an antibody fixed to the micro-particle, (b) an external magnetic field generating device for generating a magnetic field, preferably a gradient magnetic field and applying it to the solution in the test vessel to effect local concentration of the immunocomplex to a predetermined position, (c) a magnetic member for collecting the immunocomplex at the position of local concentration, and (d) a moving mechanism for achieving relative movement between the magnetic member and the test vessel.

Abstract: Poly(fluoroalkyl) sugar reagents are prepared containing a sugar such as a monosaccharide or a disaccharide to which are bonded a plurality of fluoroalkyl anchor groups capable of attaching to a fluorocarbon surface, and either a reactive group capable of covalent coupling to a biomolecule such as an enzyme or a charged group to form an ion-exchanger or a non-ionic group to give a neutral fluorosurfactant. A spacer may be between the reactive group and the sugar. The poly(fluoroalkyl) sugar reagents are strongly adsorbed onto fluorocarbon surfaces to provide supports for such applications as separation and immobilization of biomolecules such as enzymes, carrying out heterogeneous diagnostic assays, and preparation of biosensors.

Abstract: A method of separating cell nuclei from cells comprises: treating a fluid containing whole cells so as to selectively lyse the cytoplasmic membrane together with a small proportion of the nuclear membranes but leaving a large proportion of the cell nuclei intact; applying the treated fluid to a membrane whereby a mesh of DNA from the lysed nuclei is formed on the surface and captures intact cell nuclei. A device for use in the method is described.

Abstract: A method and apparatus to concentrate and purify islets of Langerhans from a tissue suspension containing islets and tissue fragments. The tissue suspension is flowed through an inclined channel such that laminar flow is established in the channel. The islets settle toward the channel bottom and are drawn out of the channel through an outlet in the channel bottom, while the remaining tissue suspension flows out a second outlet positioned higher than the islet outlet. Additional concentration or purification may be accomplished by passing the islets through the channel additional times and by centrifuging or filtering operations.

Abstract: A method is described for the use of monoclonal antibodies in a sensitive immunoassay for halogenated dioxins and dibenzofurans in industrial samples which contain impurities. Appropriate sample preparation and selective enzyme amplification of the immunoassay sensitivity permits detection of dioxin contaminants in industrial or environmental samples at concentrations in the range of a few parts per trillion.

Abstract: A fluidized bed reactor system utilizes a fluid phase, a retained fluidized primary particulate phase, and a migratory second particulate phase. The primary particulate phase is a particle such as a gel bead containing an immobilized biocatalyst. The secondary particulate phase, continuously introduced and removed in either cocurrent or countercurrent mode, acts in a secondary role such as a sorbent to continuously remove a product or by-product constituent from the fluid phase. Introduction and removal of the sorbent phase is accomplished through the use of feed screw mechanisms and multivane slurry valves.

Abstract: A present invention relates to a method for quantitatively determining sugar-alcohols characterized by passing test samples containing sugar-alcohols, proteins, and saccharides through a column filled with basic anion-exchange resins which have a protein-removing ability and a saccharide-removing ability, and then quantitaing the sugar-alcohols in the effluent out of the column, a column filled with said resins and an aqueous solution of boric acid, and a kit. Sugar-alcohols such as 1,5-anhydroglucitol and the like have been calling attention as markers for diabetes mellitus recently.

Abstract: Improvements in the existing procedures and materials for conduct of high gradient magnetic separation (HGMS) are disclosed. Superior superparamagnetic particles, optionally coated with a polysaccharide or other, usually organic, materials can be prepared in uniform compositions with homogeneous magnetizations. The coating can conveniently be conjugated to a specific binding moiety complementary to a biological material whose purification or separation is desired. In addition, plastic coated matrices which form superior magnetic gradient-intensifying supports are disclosed, along with improved methods and apparatus to conduct HGMS.

Abstract: Poly(fluoroalkyl) sugar reagents are prepared containing a sugar such as a monosaccharide or a disaccharide to which are bonded a plurality of fluoroalkyl anchor groups capable of attaching to a fluorocarbon surface, and either a reactive group capable of covalent coupling to a biomolecule such as an enzyme or a charged group to form an ion-exchanger or a non-ionic group to give a neutral fluorosurfactant. A spacer may be between the reactive group and the sugar. The poly(fluoroalkyl) sugar reagents are strongly adsorbed onto fluorocarbon surfaces to provide supports for such applications as separation and immobilization of biomolecules such as enzymes, carrying out heterogeneous diagnostic assays, and preparation of biosensors.

Abstract: A rapid, simple, sensitive and reliable method for detecting fecal carcinoembryonic antigens in stool, indicative of the presence of the colorectal cancer is described. The invention is based on the discovery that previous methods of removing coarse and gelatinous materials from a stool and liquid mixture resulted in removing a significant amount of total CEA and CEA-like substances. By not removing or destroying or altering molecules smaller than 500,000 MW, in the process of preparing the stool sample to be examined, a significant portion of CEA and CEA-like substances will remain in the filtered liquid for detection.

Abstract: A phosphazene polymer carrier is prepared that has functional groups capable of binding a biologically active substance such as an enzyme or antibody and groups which are non-reactive and hydrophilic. A bifunctional aldehyde is reacted with primary amino groups of a shaped phosphazene polymer to form side chains having aldehyde groups, an amino group-containing compound is reacted with a portion of the aldehyde groups to produce the groups that are non-reactive and hydrophilic, and aldehyde groups not reacted are capable of binding a biologically active substance. The phosphazene polymer may be crosslinked prior to reacting with the bifunctional aldehyde.

Abstract: A device for disaggregating cytological material includes a cylindrical body and a plurality of shear plates at least one of which is located at the bottom end of the cylindrical body. The shear plates have a plurality of holes or slots formed therein which disaggregate the cellular sample and permit the passage of single cells and small cell clusters into a chamber defined by the cylindrical body.

Abstract: A process for identifying proteinaceous protoxins expressed by Bacillus thuringiensis genes is disclosed. According to the process, daughter toxins are first generated by subjecting a protoxin-containing material, such as parasporal crystals of Bacillus thuringiensis, to limited proteolysis with a proteolytic enzyme in an aqueous suspension having a pH above 9.5. The daughter toxins are then separated by high performance anion-exchange liquid chromatography at a constant pH in excess of 10 in an increasing gradient of a salt, preferably sodium chloride. The gradient conditions, which are specific for the column used, are achieved by employing a series of buffers having increasing concentration of the salt and introduced at a predetermined time and rate. The procedure provides a chromatogram showing clearly identifiable peaks of toxins and permits therefore the qualitative and quantitative characterization of the original mixture and isolation of the individual toxins.

Abstract: There is disclosed a process and a device for detecting and measuring (1) the amount of enzyme present as a detecting system following a nucleic acid hybridization reaction or immunoreaction; (2) the level and activity of free enzyme in a biological sample; (3) the level of enzyme from contaminating microorganisms present in a sample; and (4) enzymes from pure culture isolates for microbial identification and antimicrobial susceptibility testing.

Abstract: A fusion protein that can function as a removable label is prepared containing a polypetide such as an enzyme and an amino acid sequence having a substrate binding region of a polysaccharidase such as cellulase that has essentially no polysaccharidase activity. By contacting the fusion protein with a .beta.-1,4 glycan matrix such as cellulose, the substrate binding region binds to the matrix to immobilize the polypeptide. The polypetide or fusion protein can be removed from the matrix with a protease capable of cleaving a specific protease cleavage site, or with a solution having a low ionic strength or a high pH. The fusion protein can be prepared by recombinant DNA technology.

Abstract: A nucleic acid is rapidly extracted from whole blood or a peripheral blood mononuclear cell (PBMC) fraction thereof. Extraction from the PBMC fraction is accomplished by heating the fraction at or near the boiling point of water for a few minutes and recovering the extracted nucleic acid. This rapid method is particularly useful for extracting DNA for the detection of genetic diseases or infectious agents, such as HIV-I. Whole blood can likewise be heated after it is mixed with a salt solution containing a polysaccharide, such as dextran. The extracted nucleic acid is then recovered from the heated mixture. Nucleic acids extracted in this way are available for amplification using a polymerase chain reaction. Where the presence of a specific gene is to be determined for diagnostic purposes, it can be extracted as described above and subjected to suitable amplification and detection steps.

Abstract: The CHLORELLA cell wall is disrupted by forming partially high- and low-pressure portions at high density in an aqueous suspension of CHLORELLA cells, instantaneously shifting the CHLORELLA cells in the aqueous suspension from a high-pressure state to a low-pressure state by interaction of the movement, dissipation and growth of these high- and low-pressure portions and the flowing of the aqueous suspension, and rupturing the CHLORELLA cells by their rapid expansion upon the shift.

Abstract: A denatured human O.sup.6 -guanine alkyltransferase is disclosed. The enzyme is prepared by a process which involves denaturing the enzyme, contacting the denatured enzyme with a monoclonal antibody specific for the denatured enzyme on a substrate to which the monoclonal antibody is bound to, so that the denatured enzyme and the monoclonal antibody form an immunocomplex and then, eluting the denatured enzyme from the substrate-bound monoclonal antibody so that the denatured human O.sup.6 -guanine alkyltransferase is obtained. The enzyme can be used to develop probes for the Mer.sup.- phenotype and these probes in turn are contemplated for use in identifying drug resistant tumors in patients.

Abstract: The lactosucrose high-content powder according to the present invention is prepared by allowing an aqueous solution containing sucrose and lactose to act on a saccharidetransferring enzyme, removing concomitant saccharides in the resultant saccharide solution containing lactosucrose to obtain a solution having a high concentration of lactosucrose, i.e. lactosucrose content of 45 w/w % or more of the sugar composition, and spraydrying the resultant solution to obtain a powder having a high concentration of lactosucrose. The powder is incorporated in an orally- or parenterally-administrable product to obtain an orally- or parenterally-administrable product which exerts a selective growth-promoting-effect on a microorganism of the genus Bifidobacterium in vivo.

Abstract: This invention provides methods and compositions for the detection of a target genetic material having a desired base sequence or gene. Also disclosed are methods and compositions for the detection of mutants. The methods and compositions are based in part upon techniques which utilize two labeled single-stranded polynucleotide segments which are complementary to different portions of the genetic material. The methods and compositions of this invention result in the formation of a double hybrid and/or multihybrid, and can furthermore be employed in an attached system, i.e., a matrix-bound entity capable of binding to a polynucleotide probe entity.

Abstract: Polymer beads are prepared containing an immobilized extractant for sorbing metal contaminants at concentrations of less than 1 mg/L in dilute aqueous solutions. A preferred polymer is polysulfone and the extractant can be a biomass material or a synthetic chemical compound sorbed into activated carbon. The polymer beads are prepared by dissolving the polymer in an organic solvent to form a solution, adding the extractant to the solution to form a mixture and injecting the mixture through a nozzle into water to form the beads.

Abstract: A fluidized bed reactor system which utilizes a fluid phase, a retained fluidized primary particulate phase, and a migratory second particulate phase. The primary particulate phase is a particle such as a gel bead containing an immobilized biocatalyst. The secondary particulate phase, continuously introduced and removed in either cocurrent or countercurrent mode, acts in a secondary role such as a sorbent to continuously remove a product or by-product constituent from the fluid phase. Introduction and removal of the sorbent phase is accomplished through the use of feed screw mechanisms and multivane slurry valves.

Abstract: A phosphazene polymer for immobilizing biologically active substances such as enzymes is prepared that does not lower activity originally possessed by the biologically active substance and does not contain a functionality which can adsorb undesired substances. The polymer has organic radicals having a functional group capable of binding a biologically active substance and organic radicals which are non-reactive and hydrophilic. The non-reactive and hydrophilic organic radicals are preferably prepared by reacting a side chain of the polymer having a primary amino group with formaldehyde or by diazotizing the primary amino group followed by hydrolysis to form a hydroxyl group. A biologically active substance immobilized on the polymer can be used to separate a substance that has affinity for the immobilized biologically active substance.

Abstract: Disclosed is a method for selecting a mutant strain belonging to the genus Streptomyces which is a hyper-producer of Streptovaricin C superior to those know heretofore. This is accomplished by first subjecting a natural strain of Streptomyces spectabilis to conditions so as to isolate organisms which are streptovaricin resistant. The streptovaricin resistant organisms thus isolated are then subjected to mutagenesis and then cultured. The colonies which are asporogenous are individually cultured in fermentation batches such that the strains take the form of pellets of varying sizes and colors. From the batch having the most heterogeneous mixture of pellets, the smallest pellet or the pellet(s) having the deepest color (usually deep red or crimson) is isolated. We have discovered that the strain of this pellet has a high likelihood of being a hyperproducer of streptovaricin. The strain of this pellet may then be subjected to fermentation conditions to produce streptovaricin.

Abstract: Polyacrylonitrile is chemically modified with HX (Cl, Br, I, CF.sub.3 SO.sub.3 H) to produce a polymer with readily replaceable X groups. The modified polyacrylonitrile is useful as an immobilization substrate for, e.g.

Abstract: A method for separation and recovery of polymeric beads from an antibiotic fermentation broth comprising suspending a mixture of the beads and any inherent mold from the fermentation in an aqueous solution having a specific gravity which is effective to cause the beads to form a discrete layer at or on the surface of the solution, separate and apart from the mold. The separate layer of the beads may then be easily removed from the liquid by a conventional physical methods.

Abstract: Method of fermentative production and isolation of Cyclosporin A by aerobic fermentation of a fungus microorganism in a medium containing mineral salts, a nitrogen-containing substrate, including using a producer strain consisting of strain Wb 6-5 of Tolypocladium inflatum, IMET 43 899 Sesquicilliopsis rosariensis G.ARNOLD, IMET 43 888, Sesquicilliopsis rosariensis G.

Abstract: A method disclosed for studying the interaction in solution of two molecules of the type such as a ligand and a receptor that are capable of reacting or binding with each other. The method comprises preparing an aliquot of a solution containing the first of the molecules. The second of the molecules is then added to the aliquot. A fluorescently labeled molecule is added to the aliquot, wherein the fluorescently labeled molecule is also capable of reacting or binding with the second of the molecules. A porous matrix that is optically transparent is immersed into the aliquot containing the two molecules being studied and the fluorescently labeled molecule, wherein the second molecule and any fluorescently labeled molecule bound thereto is sterically hindered from permeating the porous, optically transparent matrix, while any unbound fluorescently labeled molecule permeates the matrix.

Abstract: The present invention provides a process for the determination of the activity of chymotrypsin or trypsin in feces by measurement of the rate of fission of an appropriate substrate by a fecal suspension in an aqueous or aqueous-organic medium, wherein a fecal sample is suspended in the presence of a surface-active agent.The present invention also provides a reagent for carrying out this process, comprising a surface-active agent, an enzyme substrate and an aqueous salt solution, said reagent having a pH value of from 7 to 11.

Abstract: The invention provides a method for diagnosis of genetic abnormalities or other genetic conditions which can be readily automated. The method is used to determine the presence or absence of a target sequence in a sample of denatured nucleic acid and entails hybridizing the sample with a probe complementary to a diagnostic portion of the target sequence (the diagnostic probe), and with a probe complementary to a nucleotide sequence contiguous with the diagnostic portion (the contiguous probe), under conditions wherein the diagnostic probe remains bound substantially only to the sample nucleic acid containing the target sequence. The diagnostic probe and contiguous probe are then covalently attached to yield a target probe which is complementary to the target sequence, and the probes which are not attached are removed.

Abstract: Process for manufacture of D-xylose characterized by the fact that:in a first step, syrup of D-xylulose is subjected to an enzymatic isomerization in M.sub.3 providing a mixture of D-xylose and D-xylulose,in a second step, the abovesaid mixture is subjected to chromatographic treatment in M.sub.4 leading to at least two fractions of which one is highly enriched in D-xylose (fraction X.sub.1) et of which the other is highly enriched in D-xylulose (fraction X.sub.2),in a third step, the fraction X.sub.2 is recycled through a pipe P to M.sub.3,the D-xylose being recovered from the fraction X.sub.1, the latter can also be subjected directly to a hydrogenation step.