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: Disclosed is a novel derivative of human interferon-.gamma. polypeptide, a recombinant plasmid wherein a DNA fragment coding for said polypeptide is incorporated and a process for producing the derivative of human interferon-.gamma. polypeptide using a microorganism containing said plasmid.

Abstract: Interferon is produced in-vitro from whole blood, obtainable from animals or humans including blood from corpses or retroplacental blood, in a simplified non-critical process preferably carried out in a sterile system of interconnected plastic bags. The blood is stabilized and innoculated with an interferon inducer before culturing in an incubator. This results in interferon containing blood useful directly for transfusions or further processable to obtain a variety of therapeutic interferon containing preparations including plasma, serum and albumin in accordance with disclosed phases of this invention. Interferon carrying plasma is thus separated and treated further to isolate interferon carrying serum or albumin, if desired. In-vivo innoculation to produce antibodies before taking blood provides improved disease fighting characteristics in the derived product.

Abstract: Modified beta interferons containing amino acid substitutions in the beta interferon amino acids 80 to 113 are described. These modified beta interferons exhibit changes in the antiviral, cell growth regulatory or immunomodulatory activities when compared with unmodified beta interferon.

Abstract: A process is disclosed for purifying gamma interferon from various contaminants resulting from disruption of the cell in which the interferon was produced. The process provides for sequential removal of (a) nucleic acids, (b) negatively charged contaminating proteins, (c) positively charged contaminating proteins and (d) low and high molecular weight impurities.

Abstract: Human interferons containing novel cysteine substitutions and disulfide bonds are disclosed. The amino acid sequence of a first interferon is combined with the cysteine and/or disulfide pattern of a second interferon resulting in a molecule with hybrid properties.

Abstract: The human leukocyte interferon is essentially a protein featuring the foling sequence of aminoacids:CDLPQTHSLGNRRALILLAQMGRISHFSCLKDRYDFGFPQEVFDGNQFQKAQAISAF HEMIQQTFNLFSTKDSSAAWDETLLDKFYIELFQQLNDLEACVTQEVGVEEIALMNE DSILAVRKYFQRITLYLMGKKYSPCAWEVVRAEIMRSFSFSTNLQKGLRRKD.A method for producing said interferon N is bacterial cells incorporates isolation of matrix poly (A)-mRNA from induced human leukocytes, synthesis of the gene of said interferon, insertion in the vector plasmid pBR 322 under the control of a tryptophane promotor, and transformation of the resultant recombinant DNA of the E. coli bacterial cells.According to the invention, used as the interferon gene is the gene of interferon N featuring the following primary structure of DNA: ##STR1## The human leukocyte interferon features an antiviral potency and can find application in medical practice.

Abstract: A cloned human alpha-interferon Gx-1 gene, plasmids containing the human alpha-interferon Gx-1 gene, and microorganisms transformed by those plasmids are disclosed. Also disclosed is the polypeptide, alpha-interferon Gx-1.

Abstract: A peptide can be produced in a high yield by culturing a microorganism containing a recombinant DNA comprising a eukaryotic gene coding for the peptide, a vector and a promoter which is capable of producing the peptide in a medium at a temperature 10.degree. to 25.degree. C. lower than the optimum growth temperature for the microorganism.The invention is advantageously applicable to the production of peptides such as interferon.

Abstract: A process for producing HuIFN using whole human blood and a method for assaying the blood interferon (HuIFN) productivity are disclosed. The whole blood is incubated in the presence of an anticoagulant (e.g. heparin, ACD, and CPD) and a viral inducer under the conditions appropriate to accumulate a substantial amount of HuIFN. The blood HuIFN productivity determined by titrating the accumulated HuIFN with a suitable procedure (bioassay, radioimmunoassay, or enzyme-linked immunosorbent assay) is useful in clinical test to detect cancer in its early stage. The HuIFN per se is recovered, and purified prior to its prophylactic and therapeutic uses.

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 method for the treatment of essential thrombocythemia, said method comprising the administration of alpha-type interferons in an amount and for a period of time sufficient to reduce the platelet count to physiologically acceptable levels.

Abstract: Modified beta interferons containing amino acid substitutions in the beta interferon amino acids 1 to 28 are described. These modified beta interferons exhibit changes in the antiviral, cell growth regulatory or immunomodulatory activities when compared with unmodified beta interferon.

Abstract: The present invention relates to human interferon-.beta. gene derived from human chromosome, a DNA containing said gene and a DNA responsible for control of its transcription and a recombinant DNA of said DNA and a vector DNA. The gene and DNA of the present invention are introduced in eukaryotic cells and used for the production of human interferon-.beta..

Abstract: Modified beta interferons containing amino acid substitutions in the beta interferon amino acids 115 to 145 are described. These modified beta interferons exhibit changes in the antiviral, cell growth regulatory or immunomodulatory activities when compared with unmodified beta interferon.

Abstract: DNA sequences encoding for novel human lymphoblastoid-leukocyte hybrid interferons. Methods of making nucleotide sequences comprising ribosome binding sites, promoter regions, and segments of human lymphoblastoid and leukocyte interferons are described. DNA sequences for hybrid interferons are cloned and expressed in plasmid vectors. Transformed microorganisms and novel lymphoblastoid-leukocyte polypeptide interferons having biological or immunological acitivity are disclosed.

Abstract: Disclosed is a complete description of the preparation of novel, substantially pure polypeptide via recombinant DNA techniques utilizing any of an assortment of expression vectors and host cultures. The polypeptide, human immune (gamma) interferon (IFN-.gamma.), is isolated and characterized in terms of DNA and amino acid sequences, physical attributes and biological activity.

Abstract: A process for producing unique human interferon gamma and interleukin-2 by chromatographic fractionation utilizing ion exchange and metal chelate chromatography. The interferon gamma and interleukin-2 are purified from crude interferon obtained by mitogen induction of human white blood cells.

Abstract: DNA sequences encoding for novel human lymphoblastoid-leukocyte hybrid interferons. Methods of making nucleotide sequences comprising ribosome binding sites, promoter regions, and segments of human lymphoblastoid and leukocyte interferons are described. DNA sequences for hybrid interferons are cloned and expressed in plasmid vectors. Transformed microorganisms and novel lymphoblastoid-leukocyte polypeptide interferons having biological or immunological activity are disclosed.

Abstract: Adsorbents, which are suitable for the affinity chromatography of proteins, especially of interferons, and which consist of an electroneutral carrier matrix, --O--CH.sub.2 --CH(OH)--CH.sub.2 --NH- groups as the spacer and a triazine coloring substance bonded to the spacer.

Abstract: A stable human .gamma.-interferon composition is produced by adding dextran and/or hydroxyethylstarch to an aqueous human .gamma.-interferon solution freezing the resulting solution and, if desired, drying the resulting frozen solution under reduced pressure.

Abstract: The invention relates to a process for the preparation of .alpha.- and .gamma.-interferon by isolating the buffy coat fraction of blood, removing the erythrocytes, suspending the leukocytes in a suitable nutrient medium and treating with an .alpha.-interferon inducer and with a .gamma.-interferon inducer, which comprises producing .alpha.-interferon and .gamma.-interferon one after the other in the same leukocyte culture by pre-treating the suspension obtained after the separation of the leukocyte/buffy coat/fraction of blood, removal of the erythrocytes and suspending the leukocytes in a suitable nutrient medium with - or .beta.-interferon, contacting with an .alpha.-interferon inducer, separating the liquid containing the .alpha.-interferon from the cells, if desired recovering the -interferon, washing and suspending the cells in a suitable nutrient medium, treating with a mitogenic agent, separating the liquid comprising .alpha.-interferon from the cells and if desired recovering .alpha.

Abstract: Described are rapid and highly efficient procedures for the total synthesis of linear, double stranded DNA sequences in excess of about 200 base pairs in length, which sequences may comprise entire structural genes. Novel sequences are prepared from two or more DNA subunits provided with terminal regions comprising restriction endonuclease cleavage sites facilitating insertion of subunits into a selected vector for purposes of amplification during the course of the total assembly process. The total, finally-assembled assembled sequences include at least one, and preferably two or more, unique restriction endonuclease cleavage site(s) at intermediate positions along the sequence, allowing for easy excision and replacement of subunits and the correspondingly facile preparation of multiple structural analogs of polypeptides coded for by the sequences.

Abstract: A cloned human alpha-interferon Gx-1 gene, plasmids containing the human alpha-interferon Gx-1 gene, and microorganisms transformed by those plasmids are disclosed. Also disclosed is the polypeptide, alpha-interferon Gx-1.

Abstract: Monomeric human .gamma.-interferon is efficiently produced by subjecting crude human .gamma.-interferon to gel filtration in the presence of a reducing sulfur compound and a protein-denaturing agent.

Abstract: A process for producing interferon comprises culturing interferon-producing mammalian cells in a cell culture medium containing at least one compound selected from the group consisting of ascorbic acid, an ascorbic acid derivative and a vanadium compound. According to the process of the present invention, interferon can be produced in large amounts.

Abstract: A method for producing human leukocyte interferon alpha-2 resides in that there is carried out submerged cultivation of a producer strain Pseudomonas species VG-84 carrying a plasmid pVG3 with an inserted gene of human leukocyte interferon alpha-2, said strain being deposited in the collection of microorganism cultures at the USSR Antibiotics Research Institute under Reg. No. 1742; said strain being cultivated in a nutrient medium, containing the sources of carbon and nitrogen, mineral salts and growth stimulants, under aeration in the presence of antibiotics, i.e., streptomycin or tetracycline, or a mixture of both, followed by isolation and purification of the end product.

Abstract: Disclosed herein are methods and means of microbially preparing novel human hybrid leukocyte interferons, useful in the treatment of viral and neoplastic diseases, by DNA recombination of parental interferon genes, taking advantage of common restriction endonuclease cleavage sites therein and in carrier expression plasmids.

Abstract: A method for increasing the solubility of interferon in water involves admixing an amino acid selected from the group consisting of arginine, histidine, lysine, hydroxylysine, ornithine, glutamine, .gamma.-aminobutyric acid, .epsilon.-aminocaproic acid and a salt thereof with interferon.

Abstract: Disclosed is an assay procedure for detecting and quantifying interferon epsilon, a new composition of matter having selective antiviral activity on epithelial cells. The assay comprises incubating a preparation believed to contain interferon epsilon with human keratinocyte cells and with human fibroblast cells followed by a virus challenge. The presence of interferon epsilon is indicated if the preparation has antiviral activity on the keratinocytes but no detectable activity on the fibroblasts. The titer of the preparation may be determined by serially diluting it, incubating the dilution with subcultures of keratinocytes, challenging the subcultures with a virus, observing the viability of cells in the cultures, and comparing the results with a standard.

Abstract: A pharmaceutical composition for treating viral diseases, such as pasta, gargle, gel, ointment, suppository, spray, etc., containing interferon in a stable state which comprises an effective amount of human interferon, 15 to 60% by weight of a tri or higher polyhydric sugar alcohol and an organic acid buffer as stabilizers, and a conventional pharmaceutical carrier or diluent. The pH of the composition is about 3 to 6. Optionally, the composition can further contain as a stabilizer an anionic surfactant and/or albumin.

Abstract: The crystallization of human leukocyte interferon is described. In a preferred embodiment crystalline recombinant human leukocyte interferon A (IFL-rA) is prepared.

Abstract: A process for the large scale purification of human fibroblast interferon produced by human diploid fibroblasts under superinduction conditions characterized by a sequence of four chromatographic steps namely:(1) separation on CM-agarose;(2) separation on ConA-agarose;(3) separation on phenyl-agarose; and(4) elimination of ethylene glycol using CM-agarose which process yields a final overall recovery not less than 60% of the crude interferon starting material and having a specific activity not less than 1.times.10.sup.8 units per mg of protein.

Abstract: The present invention relates to processes for inserting DNA into eucaryotic cells, particularly DNA which includes a gene or genes coding for desired proteinaceous materials for which no selective criteria exist. The insertion of such DNA molecules is accomplished by cotransforming eucaryotic cells with such DNA together with a second DNA which corresponds to a gene coding for a selectable marker.This invention also concerns processes for producing proteinaceous materials such as insulin, interferon protein, growth hormone and the like which involve cotransforming eucaryotic cells with DNA which codes for these proteinaceous materials, growing the contransformed cells for production of the proteinaceous material and recovering the proteinaceous material so produced.The invention further relates to processes for inserting into eucaryotic cells a multiplicity of DNA molecules which includes genes coding for desired proteinaceous materials.

Abstract: An active fragment of human immune interferon and a method for the extraction and purification of this protein fragment is disclosed. This method permits the purification to homogenity of the active fragment of recombinant human immune interferon.

Abstract: A process and cellular system for producing soluble biological mediators, including cytokines, lymphokines, monokines and interferons, and mRNA therefor, from white blood cells is provided. The process involves treating white blood cells with a mitogenic or antigenic agent in the presence of a controlled amount of red blood cells. Increases in production of the soluble biological mediator, gamma interferon, on the order of 5 to 10 fold have been achieved by maintaining the ratio of red blood cells to white blood cells between 10 to 1 and about 50 to 1. The process is particularly useful in producing HuIFN-.gamma. with PHA-P.

Abstract: There is disclosed a filter comprising fibrin in gel form, the gel having substantially uniform pore sizes, and the filter comprising means for retaining the shape of at least one surface of the gel against deformation when contacted by a flowing medium.

Abstract: Improved vectors and methods for regulating the expression in bacteria of a eucaryotic anti-viral protein, such as mature human immune interferon, the gene for which has been cloned onto a bacterial plasmid, is disclosed. The improved vectors incorporate and method utilizes transcriptional regulatory elements derived from bacteriophage lambda and ribosome binding sites either derived from bacteriophage lambda and/or synthesized chemically.

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: 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: Methods and devices for assaying the sensitivity of of biopsied cells to therapeutic agents are disclosed. Cells are cultured in artificial organs and then contacted with a fluorogenic substrate such that living cells accumulate a characteristic amount of fluorescence. The agent is then introduced into the organ and changes in the fluorescence released by the cells serve as an indicator of the sensitivity of the cells to the agent.

Abstract: A method for the purification of human interferon by the successive treatments of a crude interferon solution with an immobilized carrier which is coupled with a blue dye and a chelating carrier which contains a chelating residue chelated with at least one metal ion selected from the group consisting of Co.sup.2+, Ni.sup.2+ and Zn.sup.2+.

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: An assay for interferon comprises two antibodies to interferon, one labelled and at least one being a monoclonal antibody. The non-labelled antibody is suitably attached to a solid support such as polystyrene beads and the labelled antibody is suitably a radiolabelled monoclonal antibody, e.g. 125 I-NK2.

Abstract: An improved process for purifying proteins, particularly proteins having molecular weights in excess of 12,000 involves novel applications of high performance liquid chromatography on a preparative scale to provide homogeneous end product in excellent yield. In an embodiment of the process, human interferon is produced as a homogeneous protein.

Abstract: A method is provided for restoring some or all of the activity of gamma-interferon which has been in contact with an acidic solution comprising the steps of: (a) placing the gamma-interferon in a solution which has a pH between about 5.5 and 9.5; and (b) incubating the solution at a temperature of between about 2.degree. C. and 8.degree. C. for a period of at least 24 hours. The method allows antibody affinity chromatography employing acid elution to be used to purify gamma-interferon, in that, the activity of the acid-eluted gamma-interferon can be essentially completely restored using the reactivation process.

Abstract: The appetite of a warm-blooded vertebrate can be regulated by administering to the vertebrate a biological active fraction of interferon in an amount effective to modulate the vertebrate's food intake or efficiency in utilizing food. The appetite of a cow can be stimulated by the oral or intravenous administration of bovine fibroblast interferon or by interferon secreted nasally by the cow in response to inoculation with a vaccinal virus strain such as that of infectious bovine rhinotracheitis virus. The appetite of swine can be enhanced by oral administration of bovine fibroblast interferon.

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: A process for the production and purification of human interferon derived from leukocyte of lymphoblastoid cell origin characterized by a sequence of steps involving chromatography of a solution of crude interferon on porous glass, utilizing three new classes of eluants for recovering the interferon bound to the porous glass, followed by cation exchange chromatography and finally chromatography on a hydrophobic sorbent material which process produces a final purified interferon in its natural form containing those protein components which are normally labile to low pH treatment along with those components which are pH stable. In one aspect, the improved method of purification also includes the optional molecular filtration of the crude interferon prior to initiating the purification process. In yet another embodiment, a new process for producingThe invention described herein was made, in part, in the course of work under grant No. NOI-HB-2920 awarded by the National Heart, Lung And Blood Institute.

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.