Abstract: Described herein are methods and genetically engineered fungal cells useful for producing target molecules containing mammalian-like complex N-glycans or containing intermediates in a mammalian glycosylation pathway.

Abstract: Optimized signal peptide coding sequences for enhanced expression and secretion of protein from a cell and related compositions and methods are described. The optimized signal peptide coding sequence encodes an mRNA that contains at least one hairpin structure immediately downstream of the initiation codon. Methods for obtaining the optimized signal peptide coding sequences and methods for enhanced expression and secretion of proteins using the optimized signal peptide coding sequences are also described.

Abstract: A process for producing an interferon alpha 5 (IFNa5) protein by expression in an IFNa5 producing Escherichia coli host cell, wherein incorporation of an extra methionine residue in the N-terminal end of the polypeptide chain is minimized as well as the generation of its oxidised species is disclosed. The IFNa5 protein can be purified by an efficient process to render a biologically active IFNa5.

Abstract: Cytokine IFN? nucleic acids from the Cynomolgus monkey are useful for expression of IFN? proteins that are functional homologs of human IFN?. The nucleic acids and proteins produced therefrom are useful in screening and safety testing of IFN?, the generation and testing of IFN? modulators and related activities.

Abstract: Novel promoters which are derived from P. pastoris pastoris which are inducible or repressible under specific growth conditions are provided. These promoters are useful for regulating the expression of a desired structural gene, e.g., a mammalian polypeptide. Particularly preferred is the use of these novel promoters to regulate gene expression in polyploidal yeast such as diploidal P. pastoris produced by mating or spheroplast fusion.

Abstract: The invention relates to variants of consensus interferon protein with improved properties, such as improved anti-viral activity, and use thereof The variants are also easier to renature after denaturant treatment. The invention also relates to the preparation method of the variants consensus interferon.

Abstract: This invention provides a recombinant super-compound interferon or an equivalent thereof with changed spatial configuration. The super-compound interferon possesses anti-viral or anti-tumor activity and, therefore, is useful in preventing and treating viral diseases and cancers. This invention also provides an artificial gene which codes for the super-compound interferon or its equivalent. Finally, this invention provides methods for producing recombinant super-compound interferon or its equivalent and various uses of said interferon.

Abstract: The present invention relates to methods of host cell transduction utilising ecotropic retroviral vector particles. The retroviral vector particle may comprise an envelope of Friend murine leukaemia virus, in particular the envelope encoded by molecular clone PVC-211 and the host cell may be engineered to recombinantly express the Reel receptor. The retroviral vector particles and methods of the invention can be used to introduce expressible polynucleotide sequences of interest into host cells with high efficiency. This results in protein production methods with higher yield (mg/L) and a reduction in manufacturing costs that could be used in a range of applications including for example, the production of therapeutic proteins, vaccines and antibodies.

Abstract: High pressure to treat aggregated interferons, particularly recombinant human interferon-?, to reduce the aggregate content of interferon material. Highly pure, soluble monomeric recombinant interferon-? is prepared in representative embodiments. Multiple strategies may be used in combination that make nonglycosylated IFN-? more amenable to high pressure treatment. It has been found that refolding yields of high pressure treatment can be significantly improved by use of a combination of strategies, including, or example a pre-treatment of the IFN-? that involves solubilizing and then precipitating the protein. This pre-treatment is particularly effective with respect to recombinant IFN-? inclusion bodies recovered from host cells such as E. coli cells. According to another strategy, refolding under high pressure is much more effective when the refolding reagent incorporating the IFN-? incorporates a zwitterionic surfactant and/or a cholate salt.

Abstract: The present invention provides a method for producing a desired protein (such as a desired heterologous protein) comprising: (a) providing a host cell comprising a first recombinant gene encoding a protein comprising the sequence of a first chaperone protein, a second recombinant gene encoding a protein comprising the sequence of a second chaperone protein and a third gene, such as a third recombinant gene, encoding a desired protein (such as a desired heterologous protein), wherein the first and second chaperones are different; and (b) culturing the host cell in a culture medium to obtain expression of the first, second and third genes.

Abstract: Methods for producing proteins and glycoproteins in Pichia pastoris that lack detectable cross binding activity to antibodies made against host cell antigens are described. In particular, methods are described wherein recombinant Pichia pastoris strains that do not display a ?-mannosyltransferase 2 activity with respect to an N-glycan or O-glycan and do not display at least one activity selected from a ?-mannosyltransferase 1, 3, and 4 activity to produce recombinant proteins and glycoproteins. These recombinant Pichia pastoris strains can produce proteins and glycoproteins that lack detectable ?-mannosidase resistant ?-mannose residues thereon and thus, lack cross binding activity to antibodies against host cell antigens. Further described are methods for producing bi-sialylated human erythropoietin in Pichia pastoris that lack detectable cross binding activity to antibodies against host cell antigens.

Abstract: A septum is positioned within a disposable vessel and defines a lower chamber and an upper chamber. The septum includes a plurality of apertures that provide fluid communication between the upper chamber and lower chamber. Compressed gas is introduced in the lower chamber to produce fine bubbles rising up throughout the vessel to produce a mixing and gasification needed for the growth of a biological culture and manufacture of a biological product in a nutrient medium. Adding a binding resin to the upper chamber allows harvesting, separation and purification of biological products in the reactor as a single unit operation.

Abstract: The present disclosure relates to a method for producing heterologous protein including: (c) providing a host cell comprising a 2 ?m-family plasmid, the plasmid comprising a gene encoding a protein comprising the sequence of a chaperone protein and a gene encoding a heterologous protein; (d) culturing the host cell in a culture medium under conditions that allow the expression of the gene encoding the chaperone protein and the gene encoding a heterologous protein; (e) purifying the thus expressed heterologous protein from the cultured host cell or the culture medium.

Abstract: The present invention provides unstructured recombinant polymers (URPs) and proteins containing one or more of the URPs. The present invention also provides microproteins, toxins and other related proteinaceous entities, as well as genetic packages displaying these entities. The present invention also provides recombinant polypeptides including vectors encoding the subject proteinaceous entities, as well as host cells comprising the vectors. The subject compositions have a variety of utilities including a range of pharmaceutical applications.

Abstract: Novel chimeric moieties that show significant efficacy against cancers are provided. In certain embodiments the chimeric moieties comprise a targeting moiety attached to an interferon. In certain embodiments, the chimeric moieties comprise fusion proteins where an antibody that specifically binds to a cancer marker is fused to interferon alpha (IFN-?) or interferon beta (IFN-?).

Abstract: Disclosed are compositions and methods for increasing the longevity of a cell culture and permitting the increased production of proteins, preferably recombinant proteins, such as antibodies, peptides, enzymes, growth factors, interleukins, interferons, hormones, and vaccines. Cells transfected with an apoptosis-inhibiting gene or vector, such as a triple mutant Bcl-2 gene, can survive longer in culture, resulting in extension of the state and yield of protein biosynthesis. Such transfected cells exhibit maximal cell densities that equal or exceed the maximal density achieved by the parent cell lines. Transfected cells can also be pre-adapted for growth in serum-free medium, greatly decreasing the time required to obtain protein production in serum-free medium. In certain methods, the pre-adapted cells can be used for protein production following transformation under serum-free conditions. The method preferably involves eukaryotic cells, more preferably mammalian cells.

Abstract: Disclosed is a method of increasing product yield per culture in a population of bound product-secreting cells in a bioreactor, the method comprising: semi-harvesting product by removing a volume of the culture medium with a first-secreted product concentration; re-feeding the bound population of product-secreting cells by adding an amount of a fresh culture medium sufficient to increase the volume of the culture medium to approximately the original volume of the culture medium; agitating the culture medium under sufficient conditions and for a sufficient time period to allow the bound population of product-secreting cells to grow and to release a second-secreted product concentration into the culture medium; and harvesting product.

Abstract: The invention provides a method for producing polypeptide protein products and nucleic acid products having reduced levels of antigenicity in an animal being treated with a biologic product. Somatic cells are isolated from an animal, transformed into pluripotent stem cells, transfected with a nucleic acid(s) of interest, and re-differentiated towards somatic cells known to be high level producers of the desired nucleic acid product. The invention can be used to derive a general cell line to treat populations, racial specific cell lines to treat ethnic groups, or patient specific cell lines to treat individuals. Additionally, the invention provides a method to allow induced pluripotent stem cells to be re-differentiated towards their somatic cell of origin so that the cells can be used to therapeutically treat an animal without resulting teratoma formation.

Abstract: The present invention discloses an improved process for the production of desired recombinant peptides from bacterial cells by using G-CSF as a novel fusion partner for their high level expression in these cells. The invention further provides an expression system comprising the fusion protein wherein the G-CSF is operatively linked to the peptide of interest via an enzymatic or chemical cleavage site which can be used to separate the fusion partner from the said peptide.

Abstract: An assembly capable of capturing and purifying expressed biological products during or at the end of a bioreaction cycle is disclosed wherein a binding resin is kept separated from the contents of the bioreactor allowing capturing, harvesting and purification of biological products in a bioreactor; the invention additionally provides means of removing undesirable metabolic products as well as provides for efficient loading of chromatography columns.

Abstract: Disclosed is an interferon-? (IFN-?) fused protein having IFN-? fused to a cytoplasmic transduction peptide (CTP). The disclosure relates to a fused protein wherein a CTP, which binds well to cell-membrane barriers and enables translocation into the liver, is genetically fused to a human IFN-?, thereby enhancing the conjugation capacity of cell membranes and antiviral activity, inhibiting CTP transport into the cell nucleus, and enhancing the translocation and settlement of the fused protein into the liver and of transduction to the liver tissue. Accordingly, it is possible to develop protein-based medicines effective for preventing or treating various liver diseases associated with viral infection at low doses.

Abstract: By culturing a methanol-assimilating bacterium which harbors a DNA construct which contains a promoter sequence that functions in the methanol-assimilating bacterium and a nucleotide sequence that encodes a polypeptide containing a signal sequence and an objective protein which is functionally connected to the promoter sequence, in a liquid medium containing methanol as a major carbon source, the bacterium is allowed to secrete the objective protein, and the secreted objective protein is recovered.

Abstract: The present invention relates to a host cell comprising a cofilin-specific small interfering RNA (siRNA) sequence. The host cell may further comprise a nucleic acid encoding a recombinant protein. The present invention also relates to a method for producing a recombinant protein by the host cell comprising a cofilin-specific small interfering RNA (siRNA) sequence.

Abstract: The present invention provides biologically active variants of human ?-2b-interferon. The variants contain carboxy terminus truncations when compared with the amino acid sequence of full-length human ?-2b-interferon. It is the novel finding of the present invention that these truncated variants have the biological activity of full-length human ?-2b-interferon. The invention encompasses these biologically active variant ?-interferons, as well as polynucleotides encoding these interferons. Expression cassettes comprising these polynucleotides and host cells comprising the expression cassettes are also provided. The invention also provides compositions comprising variant ?-interferon polypeptides and a pharmaceutically acceptable carrier.

Abstract: IFN-alpha mutants are obtained by substituting Cys for Tyr at position 85 or 86 in existing IFN-alpha. Their polyethylene glycol derivatives with high in vitro antiviral activity and prolonged in vivo half-life are also provided, wherein a polyethylene glycol moiety is covalently bound to the free Cys residue of an IFN-alpha mutant. The preparation methods of PEG derivatives of IFN-alpha mutants and medical compositions comprising the derivatives are also provided. The test results showed that the IFN-alpha mutants of the present invention are ready to prepare and have high activity; their polyethylene glycol derivatives have extended lifetime in the body and low clearance rate.

Abstract: The present invention provides methods of regulating an immunological disorder comprising administering to a subject an effective amount of (i) an autoimmune antigen in conjunction with (ii) an anti-inflammatory cytokine. Compositions including the same are also provided.

Abstract: Methods for producing modified polypeptides containing amino acid analogues are disclosed. The invention further provides purified dihydrofolate reductase polypeptides, produced by the methods of the invention, in which the methionine residues have been replaced with homoallylglycine, homoproparglycine, norvaline, norleucine, cis-crotylglycine, trans-crotylglycine, 2-aminoheptanoic acid, 2-butynylglycine and allylglycine.

Abstract: The present invention relates to novel methods of making soluble proteins having free cysteines in which a host cell is exposed to a cysteine blocking agent. The soluble proteins produced by the methods can then be modified to increase their effectiveness. Such modifications include attaching a PEG moiety to form pegylated proteins.

Abstract: This invention provides a recombinant super-compound interferon or an equivalent thereof with changed spatial configuration. The super-compound interferon possesses anti-viral or anti-tumor activity and therefore is useful to prevent and treat viral diseases and cancers. This invention also provides an artificial gene which codes for the super-compound interferon or its equivalent. Finally, this invention provides methods to produce recombinant super-compound interferon or its equivalent and various uses of said interferon.

Abstract: The present invention relates to environmental stress-inducible 557 promoter isolated from rice, a recombinant plant expression vector comprising said promoter, a method of producing a target protein by using said recombinant plant expression vector, a method of producing a transgenic plant using said recombinant plant expression vector, a transgenic plant produced by said method, a method of improving resistance of a plant to environmental stress by using said promoter, and a primer set for amplification of said promoter.

Abstract: The present invention relates to a human interferon-beta mutein. In particular, the present invention relates to the human interferon-beta mutein having one or two additional sugar chains compared to natural human interferon-beta.

Abstract: Lower eukaryotic cells such as Pichia pastoris that normally cannot use galactose as a carbon source but which have been genetically engineered according to the methods herein to use galactose as a sole source of carbon are described. The cells are genetically engineered to express several of the enzymes comprising the Leloir pathway. In particular, the cells are genetically engineered to express a galactokinase, a UDP-galactose-C4-epimerase, and a galactose-1-phosphate uridyltransferase, and optionally a galactose permease. In addition, a method is provided for improving the yield of glycoproteins that have galactose-terminated or -containing N-glycans in cells that have been genetically engineered to produce glycoproteins with N-glycans having galactose residues but which normally lack the enzymes comprising the Leloir pathway comprising transforming the cells with one or more nucleic acid molecules encoding a galactokinase, a UDP-galactose-C4-epimerase, and a galactose-1-phosphate uridyltransferase.

Abstract: The present invention relates to a process for the production of interferon beta, and to an interferon beta composition having a unique glycosylation pattern.

Abstract: The present invention relates to a method for increasing the expression of a protein in cells, preferably in eukaryotic cells, by reducing the number of RNase L cleavage sites in the coding and/or non-coding region of the nucleic acid sequence of said protein. Furthermore, it relates to nucleic acid sequences exhibiting a reduced number of RNase L cleavage sites as well as to the proteins translated from such sequences.

Abstract: The present invention relates to fusion proteins. The invention specifically relates to compositions and methods of Tf-based fusion proteins that demonstrate a high-level expression of transferrin (Tf)-based fusion proteins by inserting a helical linker between two protein domains.

Abstract: Protease resistant modified interferon-beta polypeptides, pharmaceutical compositions and methods of treatment administering the compositions are provided. The modified interferon-beta polypeptides are orally available; hence pharmaceutical compositions formulated for oral administration also are provided.

Abstract: Protease resistant modified interferon-beta polypeptides, pharmaceutical compositions and methods of treatment by administering the compositions are provided. The modified interferon-beta polypeptides are orally available; hence pharmaceutical compositions formulated for oral administration also are provided.

Abstract: The invention relates to novel polyalkylene glycol compounds and methods of using them. In particular, compounds comprising a novel polyethylene glycol conjugate are used alone, or in combination with antiviral agents to treat a viral infection, such as chronic hepatitis C.

Abstract: The present invention relates to the field of recombinant protein production in bacterial hosts. It further relates to extraction of soluble, active recombinant protein from an insoluble fraction without the use of denaturation and without the need for a refolding step. In particular, the present invention relates to a production process for obtaining high levels a soluble recombinant Type 1 interferon protein from a bacterial host.

Abstract: Compositions of modified cytokines and uses thereof generated using processes and systems for the high throughput directed evolution of peptides and proteins, particularly cytokines that act in complex biological settings, are provided. Also provided are modified cytokines formulated for oral delivery and uses thereof to treat diseases and conditions mediated by cytokines.

Abstract: This invention provides a recombinant super-compound interferon or an equivalent thereof with changed spatial configuration. The super-compound interferon possesses anti-viral or anti-tumor activity and therefore is useful to prevent and treat viral diseases and cancers. This invention also provides an artificial gene which codes for the super-compound interferon or its equivalent. Finally, this invention provides methods to produce recombinant super-compound interferon or its equivalent and various uses of said interferon.

Abstract: We provide a Chinese Hamster Ovary (CHO) cell which is capable of higher protein sialylation compared to a wild type Chinese Hamster Ovary cell, such as in the presence of functional GnT 1, in which the CHO cell is obtainable by selection with Ricinus communis agglutinin I (RCA-I).

Abstract: The present invention provides unstructured recombinant polymers (URPs) and proteins containing one or more of the URPs. The present invention also provides microproteins, toxins and other related proteinaceous entities, as well as genetic packages displaying these entities. The present invention also provides recombinant polypeptides including vectors encoding the subject proteinaceous entities, as well as host cells comprising the vectors. The subject compositions have a variety of utilities including a range of pharmaceutical applications.

Abstract: A method for the recombinant production of a polypeptide in a eukaryotic host cell modified in the citrate cycle is described, wherein the method comprises cultivating the eukaryotic host cell in a suitable medium under conditions which allow the expression of the polypeptide, wherein the content of dissolved CO2 (pCO2) in the medium is maintained at a constant value in the range of 10% to 20%.

Abstract: Methods and compositions are disclosed to engineer plastids comprising heterologous genes encoding immuno-activating domains fused to an extracellular domain (ECD) of a receptor or surface glycoprotein, a growth factor or an enzyme and produced within a subcellular organelle, such as a chloroplast. The immuno-activating domains may include those regions of a protein capable of modulating the interaction between immune effector cells via proteins containing stereoselective binding domains and specific ligands, such as the Fc regions of antibodies. The present disclosure also demonstrates the utility of plants, including green algae, for the production of complex multi-domain fusion proteins as soluble bioactive therapeutic agents.

Abstract: The present invention provides biologically active variants of human ?-2b-interferon. The variants contain carboxy terminus truncations when compared with the amino acid sequence of full-length human ?-2b-interferon. It is the novel finding of the present invention that these truncated variants have the biological activity of full-length human ?-2b-interferon. The invention encompasses these biologically active variant ?-interferons, as well as polynucleotides encoding these interferons. Expression cassettes comprising these polynucleotides and host cells comprising the expression cassettes are also provided. The invention also provides compositions comprising variant ?-interferon polypeptides and a pharmaceutically acceptable carrier.

Abstract: A transmembrane fusion protein including ubiquitin or a ubiquitin-like protein, a membrane translocation sequence linked to the C-terminus of the ubiquitin or ubiquitin-like protein, and a biologically active molecule linked to the C-terminus of the membrane translocation sequence is disclosed herein. A polynucleotide encoding the transmembrane fusion protein, a recombinant expression vector including the polynucleotide sequence, a cell transformed by the recombinant expression vector, and a method of delivering the biologically active molecule into a cell using the transmembrane fusion protein are also disclosed.

Abstract: The present invention relates to processes for the production of peptides, and the peptides produced accordingly. Peptides produced according to the invention may be produced more efficiently than peptides produced according to prior art processes. The production process of the invention may lead to advantages in yield, purity, and/or price. Methods of marketing peptides are also disclosed.

Abstract: The present invention relates generally to glutamine-free cell culture media supplemented with arginine. The invention further concerns the production of recombinant proteins, such as antibodies, in arginine-supplemented glutamine-free mammalian cell culture.

Abstract: The present invention relates to a novel process for the production of interferon-beta in improved yields by fermentation.