Abstract: A refractile material containing a heterologous protein is recovered from a host microorganism cell culture transformed to produce the protein. One recovery process involves disrupting the cell wall and membrane of the host cell, removing greater than 99% by weight of the salts from the disruptate, redisrupting the desalted disruptate, adding a material to the disruptate to create a density or viscosity gradient in the liquid within the disruptate, and separating the refractile material from the cellular debris by high-speed centrifugation. Another version of such a recovery process comprises the further steps of solubilizing the refractile material under reducing conditions, organically extracting the solubilized refractile material, and isolating said refractile material from the extractant.Preferably the protein is recombinant IL-2 or IFN-.beta. and the salt removal step is carried out by diafiltration.

Abstract: A pharmaceutical composition is prepared wherein a biologically active conjugated protein which is .beta.-interferon, interleukin-2, or an immunotoxin is dissolved in an aqueous carrier medium without the presence of a solubilizing agent. The unconjugated protein, which is poorly or not at all water-soluble at pH 6-8 without such solubilizing agent, is selectively conjugated to at least one heparin fragment having a terminal 2,5-anhydro-D-mannose residue which has an aldehyde not involved in intramolecular hemiacetal formation.

Abstract: A modified IFN-.beta. is provided wherein the cysteine residue at position 17 is deleted and serine is substituted therefor. DNA sequences coding for the modified protein, nucleotide primers used for the mutagenesis, appropriate cloning vectors, host organisms transformed with the vectors, methods for the production and use of the modified IFN-.beta. (IFN-.beta..sub.ser17) are also provided. The specific activity of IFN-.beta..sub.ser17 is found to be substantially the same as that of native IFN-.beta..

Abstract: Method for preparing peptides free of undesired amino acids or amino acid sequences employing site specific receptors and proteases to cleave unprotected enzymatically hydrolyzable bonds.

Abstract: A method of rapid determination of the isotype class for a panel of monoclonal antibodies is described. The assay comprises adsorbing on a solid support medium antibodies directed to specific immunoglobulin heavy and light chains. Once such isotype-specific antibodies are bound to the nitrocellulose paper, the treated strips can be incubated with the monoclonal antibody of interest. Upon formation of a complex between the specific iso-type antisera and the monoclonal antibody, the complex is visualized by reaction with a chromogenic substance. In the preferred embodiment of the invention, the treated nitrocellulose strips are stored in kit form. Using these prepared strips, the isotyping assay can be performed in less than two hours with a minimum of technical manipulation and expenditure of reagents.

Abstract: A method and a cloning vector are described for the controlled accumulation of cloned heterologous gene products in Bacillus subtilis. The cloning vector is capable of being replicated in B. subtilis and includes the heterologous gene located and oriented such as to be under the control of an operator, promoter, and ribosomal binding site sequence. The gene codes for a protein which is under the control of a transport mechanism by which the protein is secreted by the B. subtilis. The gene product is recovered from the growth medium for the B. subtilis. The cloning vector is also capable of similar use in other bacteria such as E. coli.

Abstract: The invention discloses a method for producing modified signal peptides sequences derived from wild-type signal peptide sequences of the type that are capable of forming membrane-bound lipoproteins. Modified signal peptide sequences produced by the method of the invention are useful for increasing the secretion of heterologous gene products produced by transformed host organisms. The invention further discloses a method for producing recombinant DNA sequences in vivo.

Abstract: Two types of convenient portable control cassettes for the expression of protein encoding sequences in procaryotes are disclosed. Both cassettes comprise the P.sub.L promoter from lambda phage, which is controllable by means of a temperature sensitive repressor, operably linked to the ribosome binding site for N-gene (N.sub.RBS). In one embodiment, this cassette is bordered by restriction sites upstream of the P.sub.L promoter and immediately downstream from the N.sub.RBS permitting the insertion of a desired sequence containing its own start codon downstream from the cassette. The other embodiment contains an ATG start codon within the cassette and has a restriction site immediately 3' of the start codon.

Abstract: A method of producing a non-heme haloperoxidase which is substantially resistant to inactivation, at room temperature, in up to 0.3M H.sub.2 O.sub.2 for up to 25 hours, and up to 0.5 mM HOCl for up to two minutes. One such haloperoxidase, isolated from Curvularia inaequalis, contains about 2 gram atoms of zinc per molecule. A halogenation reaction employing the enzyme can be performed at H.sub.2 O.sub.2 and hypohalous acid concentrations which produce rapid inactivation of heme-containing haloperoxidases.

Abstract: A halogenation method using a haloperoxidase obtained from a fungus selected from the dematiaceous hyphomycetes. The enzyme has an optimum activity above about pH 5.0, and can oxidize chloride, bromide, or iodide ions.

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 moiety and L is biotin.

Abstract: Polypeptides that are synthesized or expressed directly in microorganisms, include an amino acid sequence that is homologous to at least a portion of gp70 envelope protein of FELV, and are immunogens that elicit or prime a humoral response in cats and immunize cats against FeLV infection. The microbially produced polypeptides are exemplified by a group of E. coli-expressed fusion proteins that comprise a portion of the trp leader peptide and a portion of the trp E protein fused to various portions of a subgroup B gp70 envelope protein. The synthesized polypeptides are exemplified by polypeptides that are homologous to segments of the hydrophilic domains of the gp70 protein.

Abstract: An improved blind label is disclosed which includes a combination of open text and blinded text arrayed upon adhesive-backed label stock in an arrangement in which the label is divided into two sections joined by a tearable separation zone. In this improved label the first section is permanently affixable to the container to be labeled by its adhesive. The second section, carrying at least a portion of the blinded text, is strippably affixable to the container or to the first section by means of its adhesive backing. In certain embodiments, the blinding is accomplished by means of an opaque overcovering strippably adhered over the text with tamper-evident metallic layers being preferred overcoverings. An automatic labeling machine-compatible blind-label feedstock is also disclosed.

Abstract: At least two antigens in a sample may be detected using an immunometric dual sandwich assay containing an effective amount of at least one monoclonal antibody against each antigen, which antibodies are separately conjugated with the same or different signal moieties as labels, and an effective amount of at least one unlabeled monoclonal antibody against each antigen which unlabeled antibodies are immobilized on a single support. Preferably the antibodies are all products of different cell lines and the antigens are prostatic acid phosphatase and prostate antigen.

Abstract: Substantially pure, intracellularly produced, soluble recombinant ricin toxin A (RTA) is recovered from transformed cells by disrupting the cell membrane, removing insoluble cell membrane materials from the disruptate, adjusting the pH of the cell membrane material-free solution to 6 to 6.5 and the conductivity to 1.25 to 1.75 millisiemens, passing the adjusted solution through a bed of SP-cellulose cation exchanger, and eluting the substantially pure RTA from the bed.

Abstract: A method for controlling weight by suppressing the normal metabolism of adipose tissue is disclosed. Administration of tumor necrosis factor (TNF) or a pharmaceutical composition containing it results in suppression of anabolism of adipose cells.

Abstract: In a method for detecting the presence or absence of a specific restriction site in a nucleic acid sequence an oligonucleotide probe complementary to one strand of the nucleic acid sequence spanning said restriction site is synthesized. The probe is labeled at the end nearer the restriction site. The nucleic acid is hybridized to the probe and a blocking oligomer may be added, if necessary, to prevent non-specific binding of the probe. Subsequent digestion with a restriction enzyme cleaves those oligomers that have hybridized to the nucleic acid and reformed the restriction site. The resulting cut and uncut labeled oligomers are separated and detected based on the type of probe label.The described method may be used to detect sickle cell anemia.

Abstract: The present invention is directed to a process for amplifying any desired specific nucleic acid sequence contained in a nucleic acid or mixture thereof. The process comprises treating separate complementary strands of the nucleic acid with a molar excess of two oligonucleotide primers, and extending the primers to form complementary primer extension products which act as templates for synthesizing the desired nucleic acid sequence. The steps of the reaction may be carried out stepwise or simultaneously and can be repeated as often as desired.

Abstract: The present invention is directed to a process for amplifying and detecting any target nucleic acid sequence contained in a nucleic acid or mixture thereof. The process comprises treating separate complementary strands of the nucleic acid with a molar excess of two oligonucleotide primers, extending the primers to form complementary primer extension products which act as templates for synthesizing the desired nucleic acid sequence, and detecting the sequence so amplified. The steps of the reaction may be carried out stepwise or simultaneously and can be repeated as often as desired.In addition, a specific nucleic acid sequence may be cloned into a vector by using primers to amplify the sequence, which contain restriction sites on their non-complementary ends, and a nucleic acid fragment may be prepared from an existing shorter fragment using the amplification process.

Abstract: A machine for transferring liquids to and from the wells of assay trays in a controlled, automated manner and a solid phase assay tray for use with the machine. The machine includes a horizontally translatable table (15) that holds the tray (46), a plurality of liquid dispensing manifolds (54) for dispensing liquids into the tray wells (50) and an aspirating manifold (65) for aspirating liquid from the well. The dispensing and aspirating manifolds are mounted on a vertically translatable head (16) above the table. Each dispensing manifold is equipped with a row of dispensing tubes (56) and is connected via a pump (58) to a liquid container (62). The aspirating manifold is equipped with a row of aspirating tube (66) and is connected via a pump (69) to a waste liquid receptacle (73). A microprocessor (85) controls the movements of the table and manifolds and operates the pumps.