Abstract: Human tumor necrosis factor (TNF) has been prepared using recombinant methods. A human promyelocytic leukemia cell line has been induced using an improved induction procedure, and the TNF purified to homogeneity. Methods, vectors, and cells useful in obtaining human TNF in practical amounts are disclosed. Muteins having N-terminal deletions, which muteins have superior biological activity, are also disclosed.

Abstract: Anti-Pseudomonas antibodies which bind to exotoxin A from P. aeruginosa are prepared from hybrid cell lines. The antibodies may be of any isotype. These antibodies may be used to treat infections caused by P. aeruginosa.

Abstract: Human Tumor necrosis factor (TNF) has been prepared using recombinant methods. A human promyelocytic leukemia cell line has been induced using an improved induction procedure, and the TNF purified to homogeneity. Methods, vectors, and cells useful in obtaining human TNF in practical amounts are disclosed. Muteins having N-terminal deletions, which muteins have superior biological activity, are also disclosed.

Abstract: An improved method for purifying TNF, especially recombinantly produced TNF in bacteria, is disclosed. The purification employs a hydrophobic support in a chromatographic column which is then developed, preferably by HPLC. Preliminary purifications using anion exchange resins are also helpful.

Abstract: The invention is a plasmid vector in which a first DNA sequence expressionable for a selected gene product is ligated to a positive retroregulatory element in a relationship thereto whereby expression of the selected gene product is enhanced. The first DNA sequence is furthermore operably linked to a second and third DNA sequence operably linked to one another which are respectively the P.sub.L promoter and a DNA sequence corresponding to an N gene ribosome binding site. Microorganisms transformed with the plasmid are also claimed.

Abstract: A method for improving the yield of heterologous protein such as IFN-.alpha., IFN-.beta., and IL-2, produced by recombinant bacteria by supplementing the nutrient medium in which the bacteria are grown with a water soluble alcohol and/or amino acid mixture during the terminal phase of the cultivation.

Abstract: Catalase and pyranose-2-oxidase are stabilized according to the invention by means of chemical treatment which render the enzymes resistant to thermal inactivation or inactivation of glucosone or both.Stabilized catalase crosslinked with diimido esters, such as dimethyl suberimidate and dimethyl adipimidate are claimed.The method for stabilizing catalase against thermal inactivation or glucosone inactivation or both comprising gradually adding a crosslinking agent to a catalase solution and maintaining pH and temperature control is claimed.Stabilized pyranose-2-oxidase amidinated with an amidinating agent such as ethyl acetimidate is claimed.A method for stabilizing catalase against thermal inactivation comprising gradually adding an amidinating agent to a pyranose-2-oxidase solution preferably with pH control is claimed.An improved process for producing glucosone from glucose using stabilized catalase or pyranose-2-oxidase or both according to the invention is also claimed.

Abstract: An apparatus and process for storing beads in a liquid medium and for draining and dispensing said beads essentially in the absence of said liquid medium is described. In one embodiment, the apparatus is designed so that the liquid medium in which the beads are stored can be captured in a separate compartment from the bead-containing compartment and the beads can be delivered without the liquid medium when the apparatus in inverted. This apparatus is designed so that the liquid medium will join with the beads for storage in the bead-containing compartment when the apparatus is upright. In another embodiment, the apparatus includes a bottle with a removable, threaded inner container which is used to drain the liquid medium from the beads prior to attachment to a separate bead dispenser.

Abstract: DNA plasmids are described which are selected mutants in which an altered repressor gene leads to high copy number replication. Elements in the plasmids are modified in such a way that readthrough expression of heterologous DNA inserted in the plasmid will not continue into the replication primer strand. Deletions resulting from interference with replication primer strand transcription are thereby avoided.

Abstract: A 6-thioguanine-resistant subvariant of the EBV-transformed human lymphoblastoid B cell line WI-L2 is described. The subvariant line, designated LTR228, fuses efficiently with human cells. Human.times.human hybridomas derived from LTR228 that produce monoclonal antibodies against tetanus toxin and blood group A are exemplified.

Abstract: Monoclonal antibodies having conformation-dependent specificity for native dsDNA, as exemplified by the IgM antibody produced by murine hybridoma ATCC No HB 8329. These antibodies are used to detect DNA duplex formation in DNA hybridization tests.

Abstract: Nucleic acids may be labeled by intercalating the alkylating intercalation moiety of a labeling reagent into a partially double-stranded nucleic acid to form a complex and activating the complex to cause covalent bonding between the reagent and the nucleic acid. Preferably, the labeled nucleic acid is a hybridization probe for detecting nucleic acid sequences capable of hybridizing with a hybridizing region of the nucleic acid. Also preferably the label moiety is non-radioactive. The labeling reagent is of the formula:[A--[B--Lwhere A is an alkylating intercalation moiety, B is a divalent organic moiety of the formula: ##STR1## where Y is O, NH or N--CHO, x is a number from 1 to 4, y is a number from 2 to 4, and L is a monovalent label moiety, wherein B is exclusive of any portion of the intercalation and label moieties.Preferably A is a 4-methylene-substituted psoralen moiety, and most preferably A is a 4'-methylene-substituted-4,5',8-trimethylpsoralen moeity and L is biotin.

Abstract: A sterile, stable lyophilized formulation of selectively oxidized microbially produced recombinant IL-2 in which the recombinant IL-2 is admixed with a water soluble carrier such as mannitol that provides bulk, and a sufficient amount of sodium dodecyl sulfate to ensure the solubility of the recombinant IL-2 in water. The formulation is suitable for reconstitution in aqueous injections for parenteral administration and it is stable and well tolerated in human patients. FIG. 1 illustrates a preferred method of purifying recombinant IL-2 suitable for use in preparing the formulations of the present invention.

Abstract: Muteins of biologically active proteins such as IFN-.beta. and IL-2 in which cysteine residues that are not essential to biological activity have been deleted or replaced with other amino acids to eliminate sites for intermolecular crosslinking or incorrect intramolecular disulfide bridge formation. These muteins are made via bacterial expression of mutant genes that encode the muteins that have been synthesized from the genes for the parent proteins by oligonucleotide-directed mutagenesis.

Abstract: A method is provided for the production of vicinal heterogeneous dihalogenated products from alkenes and alkynes. The alkene or alkyne is reacted in an aqueous solution of salts of two different halides, an oxidizing agent and an halogenating enzyme. Products provided by the method include novel 2,3-heterogeneously dihalogenated-1,4-butanediols.

Abstract: An improved liquid handling device used for transferring a selected quantity of liquid from one receptacle to another. The device includes a meniscus tracking feature which minimizes mixing in a liquid sample and contact between the sample and a sample-handling pipette during a liquid-transfer operation. Further included is a volume-correcting feature for improving the accuracy of volume withdrawn into or dispensed from the pipette during a liquid handling operation.

Abstract: A labeling reagent of the formula:[A][B]Lis prepared where A is an alkylating intercalation moiety, B is a divalent organic spacer arm moiety with a straight chain of at least two carbon atoms, and L is a monovalent label moiety capable of producing a detectable signal, e.g., a signal detectable by spectroscopic, photochemical, chemical, immunochemical or biochemical means. Preferably A is a 4'-methylene-substituted psoralen moiety, and most preferably A is a 4'-methylene-substituted 4,5',8-trimethylpsoralen moiety.This reagent may be used to label nucleic acids, preferably DNA, by intercalating the alkylating intercalation moiety of the reagent into an at least partially double-stranded nucleic acid to form a complex and activating the complex to cause covalent bonding between the reagent and the nucleic acid. Preferably, the labeled nucleic acid is a hybridization probe for detecting nucleic acid sequences capable of hybridizing with a hydridizing region of the nucleic acid.

Abstract: HLA typing based on restriction length polymorphism is carried out by: digesting an individual's HLA DNA with a restriction endonuclease that produces a polymorphic digestion pattern with HLA DNA; subjecting the digest to genomic blotting using a labeled cDNA hybridization probe that is complementary to an HLA DNA sequence involved in the polymorphism; and comparing the resulting genomic blotting pattern with a standard. This technique may be adapted to make paternity or transplant or transfusion compatibility determinations or to make disease association correlations to diagnose diseases or predict susceptibility to diseases. Locus specific cDNA hybridization probes, particularly probes for genes of Class II loci (D and DR loci), for use in the typing procedure are described.

Abstract: Recombinant plasmids are described that carry the structural gene for human proinsulin in expressible form. The human proinsulin structural gene is fused to a functional transport sequence. Host organisms transformed by the recombinant plasmids express a preproinsulin in vivo, and is transported across the host organism's cell wall. Following its recovery, in vitro methods are used to convert the human proinsulin to human insulin.