Abstract: Reduced cysteine-containing proteins consisting of recombinant IFN-.beta., IL-2 or muteins thereof may be oxidized selectively so that the recombinant proteins have essentially the same disulfide bridging and biological activity as their native counterparts. The oxidized product is substantially free of unwanted side products and contains a minimal amount of intermolecular oligomers. The oxidation takes place in an aqueous medium containing a solubilizing agent at a pH of about 5.5 to 9, preferably at a pH of about 7. The reaction is initiated by addition of at least an effective amount of an oxidation promoter containing a Cu.sup.+2 cation such as CuCl.sub.2 or o-phenanthroline/Cu.sup.+2 complex in the presence of air.

Abstract: A process for recovering microbially produced IL-2 in a highly pure form from the cellular material of the microorganisms that produced it comprising: disrupting the cell membranes of the microorganisms; extracting the disruptate with a chaotropic agent, such as urea, that selectively extracts microbial proteins from the cellular material; solubilizing the IL-2 in the solid phase of the extraction mixture with an aqueous solution of a solubilizing agent, such as SDS, containing a reducing agent; and separating the IL-2 from the resulting solution by an optional extraction with 2-butanol or 2-methyl-2-butanol followed by gel filtration chromatography, oxidizing the IL-2 and purifying the oxidized IL-2 by RP-HPLC.

Abstract: Methods for producing substantially pure pyranosone from pyranose, especially glucosone from glucose. A pyranose-2-oxidase enzyme which is free of substantially all glucosone-utilizing enzyme contaminants which are measurably active at a selected pH between about 4.4 and 7.0 is reacted in solution with a pyranose and oxygen to produce the corresponding pyranosone. Hydrogen peroxide co-produced in the reaction is maintained at a concentration below about 10.sup.-3 M, by substantially pure catalase. The reaction is carried out at the selected pH. The methods and reagents are also applicable to producing xylosone from xylose, 5-ketofructose from sorbose, and mixtures of 2-ketogluconic and isoascorbic acid from delta-gluconolactone.

Abstract: A P. obtusus strain AU124 having the identifying characteristics of NRRL No. 15595, the strain produces high levels of pyranose-2-oxidase.

Abstract: A substantially pure pyranose-2-oxidase preparation obtained from P. obtusus, and preferably from a high-producer strain thereof. The preparation is substantially free of glucosone-utilizing enzyme contaminants having measurable activity at a pH between about 4.4 and 7.0.

Abstract: New multiclass hybrid interferon polypeptides, their corresponding encoding recombinant DNA molecules and transformed hosts which produce the new interferons are described. The amino acid sequences of these hybrids include at least two different subsequences, one of which has substantial homology with a portion of a first class of interferon (e.g., HuIFN-.alpha.) and the other which has substantial homology with a portion of a second class of interferon e.g., HuIFN-.beta.). Data indicates the interferon activity of .alpha.-.beta. hybrids may be substantially restricted to either cell growth regulatory activity or antiviral activity.

Abstract: A substantially pure pyranose-2-oxidase preparation obtained from C. versicolor or L. betulinus. The preparation is substantially free of glucosone-utilizing enzyme contaminants having measurable activity at a pH between about 4.4 and 7.0.

Abstract: A process for producing wax esters having the general structural formula RCO.sub.2 R', where R and R' are radicals selected from the group consisting of CH.sub.3 (CH.sub.2).sub.a CH.dbd.CH(CH.sub.2).sub.b - and CH.sub.3 (CH.sub.2).sub.c - where a+b=12-14 for R and 13-15 for R' and where c=14-16 for R and 15-17 for R'. The method includes aerobically incubating a culture of microorganisms of the genus Acinetobacter species HO1-N, in an aqueous mineral salts solution containing ethanol as a primary food source. Incubation is performed at a temperature known to produce a desired percentage of diene, monoene and saturated R and R' moieties in the mixture.

Abstract: An automatic liquid transfer system includes a horizontally translatable table and a pipette that is translatable both vertically and horizontally in a direction transverse to the axis of translation of the table. The table accommodates one or more titer trays having a multiplicity of receptacles to be filled, or holding liquid samples to be diluted, transferred, or aspirated, and a rack housing plural rows of disposable tips. Between each cycle in a liquid transfer process, a fresh tip is picked up by the pipette and used to transfer liquid in a sterile manner from a sample or diluent source to a well in the titer tray, or from one well to a different well anywhere on the one tray or to a receptacle in a different tray, where it is mixed with diluent. Thereafter, the tip is discharged back into the rack to maintain sterile conditions during the process and a new tip is installed.

Abstract: A method is described for constructing double-stranded DNA using messenger RNA templates. DNA strands complementary to the templates are formed using a non-specific 3' hydroxy terminus primer. After separation, the DNA single strands are disabled from self priming by blocking the free 3' hydroxy. Primer segments are then provided complementary to a portion of the amino terminus-coding end of the desired single strands. These primer segments are then extended with a suitable polymerase to complete the complementary strands.

Abstract: Apparatus for accurately transferring liquids for filling or diluting microliter quantities of regents, analytes or cells for bioassays and the like include a vessel having a plurality of troughs for supplying an array of pipettes to effect the transfer. The vessel is automatically indexable on a horizontal table to bring a selected trough into register with vertical movement of the pipette tips. The table is then indexed to bring the tips into alignment with a plurality of microtiter tray wells. In a preferred form the troughs are of different widths to supply differing quantities of liquid to the pipettes for transfer.

Abstract: A method is described for the manufacture of commercially useful alpha-halo and alpha-dihalo ketones and aldehydes from alkynes by enzymatic reaction. The alkyne is acted upon in a reaction mixture comprising a halogenating enzyme, an oxidizing agent, and a halide ion source.

Abstract: Method of oxidizing reduced cysteine-containing microbially produced synthetic proteins, such as synthetic IFN-.beta. or synthetic IL-2, in a controlled manner so that the synthetic proteins have the same disulfide bridging as their native counterparts. The oxidation employs o-iodosobenzoate as oxidizing agent and is carried out in an aqueous medium at a pH at least about one-half pH unit less than the pK.sub.a of the cysteines to be oxidized, a synthetic protein concentration of less than about 5 mg/ml, and an oxidizing agent:protein mol ratio that is at least stoichiometric, provided that the oxidizing agent is in excess in the terminal portion of the reaction.

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 described for the manufacture of aldehydes from primary alcohols by enzymatic reaction. The aldehydes are produced by contacting a primary alcohol with a reaction mixture consisting of chloroperoxidase and hydrogen peroxide.

Abstract: A process for bacterially producing heterologous polypeptides, particularly those such as human IFN-.beta. that inhibit bacterial growth, in which bacteria that have been transformed to express the heterologous polypeptide under the control of a trp promoter-operator are cultivated in a known volume of medium containing an excess of a preferred carbon source such as glucose and a predetermined amount of tryptophan that corresponds approximately to the amount of tryptophan contained in the bacteria in the volume of medium at a predetermined elevated cellular density, whereby expression of the heterologous polypeptide is substantially repressed until the bacteria grow to approximately the predetermined elevated cellular density and is thereafter automatically derepressed to permit expression of the heterologous polypeptide.

Abstract: An improved process for the isolation and purification of HIFNs is disclosed wherein a partially purified preparation of the HIFN is sequentially passed through an antibody affinity column and a reversed-phase high performance liquid chromatographic column. Organic solvents used during the elution are extracted and the protein concentrated for subsequent use.

Abstract: An automatic liquid transfer system includes a horizontally translatable table and a vertically translatable set of pipettes. The table accommodates a titer tray having a multiplicity of receptacles to be filled, or holding liquid samples to be diluted, and a rack housing plural rows of disposable tips. During each cycle in a serial dilution process, a fresh set of tips are picked up by the pipettes and used to transfer liquid in a sterile manner from a sample or diluent source to a row of wells in the titer tray, or from one row to a succeeding row of wells where it is mixed with diluent. Thereafter, the tips are discharged back into the rack to maintain sterile conditions during the process. A sensor is provided on the machine to detect whether all of the tips in each set are disengaged and another set successfully picked up.

Abstract: Chimeric plasmids capable of transforming bacteria and yeast are described. The plasmids carry the Cm (chloramphenicol resistance) gene and the Tc (tetracycline resistance) gene as selectable markers. The Cm gene allows the plasmids to be selected for in wild-type strains of the yeast Saccharomyces cerevisiae. The Tc gene allows heterologous genes cloned into the plasmids to be selected for in Escherichia coli bacteria.

Abstract: A composition of matter exhibiting a synergistic cytotoxic effect in combination therapy of certain breast cancer and myeloma cell lines is prepared by combining synergistically effective amounts of 5-fluorouracil and human recombinant beta-interferon, optionally together with a pharmaceutically effective diluent or carrier. Preferably the recombinant beta-interferon is a mutein in which the cysteine residue at position 17 of native beta-interferon is replaced by a serine residue.