Abstract: This invention provides a set of methods for modulating protein spatial configuration. First, select the amino-acid codon for encoding the target protein according to host codon usage. Second, choose combinations which can modulate the spatial configuration and construct into different vectors which can transfect a series of hosts. Third, choose the vector promoter by monitoring a combination of base pairs after combining the code sequence of the promoter and the target protein. Finally, choose the appropriate expression host to express the target protein, refold and purify, measure the activity and spatial configuration.

Abstract: The present invention relates to novel expression cassettes and vectors for efficiently producing authentic recombinant human proteins from stable cultures of novel human cell lines, the authentic recombinant proteins produced therefrom, and antibodies raised against those authentic recombinant proteins.

Abstract: The current invention reports a method for the purification of a not-glycosylated, heterologous polypeptide, which has been recombinantly produced in a prokaryotic cell, wherein the method comprises three chromatography steps of which the first chromatography step selected from i) hydrophobic charge induction chromatography, or ii) hydrophobic interaction chromatography, or iii) affinity chromatography, or iv) ion exchange chromatography, the second chromatography step is selected from i) anion exchange chromatography, or ii) cation exchange chromatography, or iii) hydroxylapatite chromatography, or iv) hydrophobic interaction chromatography, and the a third chromatography step is selected from i) hydrophobic charge induction chromatography, or ii) anion exchange chromatography, or iii) cation exchange chromatography, or iv) hydrophobic interaction chromatography, whereby the first chromatography step is an affinity chromatography in case of polypeptides capable of interacting with metal ligands, the second c

Abstract: Cell lines having genetically modified glycosylation pathways that allow them to carry out a sequence of enzymatic reactions, which mimic the processing of glycoproteins in humans, have been developed. Recombinant proteins expressed in these engineered hosts yield glycoproteins more similar, if not substantially identical, to their human counterparts. The lower eukaryotes, which ordinarily produce high-mannose containing N-glycans, including unicellular and multicellular fungi are modified to produce N-glycans such as Man5GlcNAc2 or other structures along human glycosylation pathways.

Abstract: The invention relates to an expression vector comprising the promoter of the mCMV-IE2 gene, or a functional expression promoting fragment thereof, and/or an enhancer of the mCMV-IE2 gene, or a functional expression enhancing fragment thereof, wherein the expression vector does not contain any complete gene of the mCMV.

Abstract: This application relates to recombinant human interferon-like proteins. In one embodiment a recombinant protein created by gene shuffling technology is described having enhanced anti-viral and anti-proliferative activities in comparison to naturally occurring human interferon alpha 2b (HuIFN-?2b). The invention encompasses a polynucleotide encoding the protein and recombinant vectors and host cells comprising the polynucleotide. Preferably the polynucleotide is selected from the group of polynucleotides each having a sequence at least 93% identical to SEQ ID: No. 1 and the protein is selected from the group of proteins each having an amino acid sequence at least 85% identical to SEQ ID No: 2. The proteins and compositions comprising the proteins can be used for treatment of conditions responsive to interferon therapy, such as viral diseases and cancer.

Abstract: This application relates to recombinant human interferon-like proteins. In one embodiment a recombinant protein created by gene shuffling technology is described having enhanced anti-viral and anti-proliferative activities in comparison to naturally occurring human interferon alpha 2b (HuIFN-?2b). The invention encompasses a polynucleotide encoding the protein and recombinant vectors and host cells comprising the polynucleotide. Preferably the polynucleotide is selected from the group of polynucleotides each having a sequence at least 93% identical to SEQ ID: No. 1 and the protein is selected from the group of proteins each having an amino acid sequence at least 85% identical to SEQ ID No: 2. The proteins and compositions comprising the proteins can be used for treatment of conditions responsive to interferon therapy, such as viral diseases and cancer.

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: The present invention relates to human interferon gamma variants with improved thermostability, to a nucleic acid encoding said variants, to a pharmaceutical composition containing them, and to their use for the treatment of a viral infection and of cancer.

Abstract: The present invention provides Interferon-Like (IFN-L) polypeptides and nucleic acid molecules encoding the same. The invention also provides selective binding agents, vectors, host cells, and methods for producing IFN-L polypeptides. The invention further provides pharmaceutical compositions and methods for the diagnosis, treatment, amelioration, and/or prevention of diseases, disorders, and conditions associated with IFN-L polypeptides.

Abstract: The invention provides compositions and methods of identifying, modifying and producing modified target molecules, including therapeutic molecules by modification with non-natural amino acids. Certain aspects of the invention include methods of adding a chemical moiety to a target molecule, and the compositions resulting therefrom. Certain aspects of the invention also relate to kits for identifying, modifying and producing modified target molecules described herein.

Abstract: A method is provided for introducing a GAG binding site into a protein comprising the steps: identifying a region in a protein which is not essential for structure maintenance introducing at least one basic amino acid into said site and/or deleting at least one bulky and/or acidic amino acid in said site, whereby said GAG binding site has a GAG binding affinity of Kd?10 ?M, preferably ?1 ?M, still preferred ?0.1 ?M, as well as modified GAG binding proteins.

Abstract: The growth hormone supergene family comprises greater than 20 structurally related cytokines and growth factors. A general method is provided for creating site-specific, biologically active conjugates of these proteins. The method involves adding cysteine residues to non-essential regions of the proteins or substituting cysteine residues for non-essential amino acids in the proteins using site-directed mutagenesis and then covalently coupling a cysteine-reactive polymer or other type of cysteine-reactive moiety to the proteins via the added cysteine residue. Disclosed herein are preferred sites for adding cysteine residues or introducing cysteine substitutions into the proteins, and the proteins and protein derivatives produced thereby.

Abstract: The present disclosure provides isolated Interferon? nucleic acids and polypeptides. The disclosure also provides antibodies which specifically recognize the subject Interferon? polypeptides, expression vectors containing the subject nucleic acids, and host cells expressing the subject polypeptides. In addition, methods of treatment using Interferon? are provided.

Abstract: Method for heterologous protein production in plant cell plastids comprising introducing into plant cells nucleic acid components that encode heterologous proteins under the control of promoters operative in plastids, vectors, host cells, plants and uses thereof.

Abstract: The present invention relates to mutant polypeptides of the beta chain of the type I IFN receptor (IFNAR2 mutant) with enhanced affinity for IFN? as compared to the wild type protein for prolonging the effect of IFN? in vivo.

Abstract: Expression cassettes are provided comprising a promoter operably linked to a nucleic acid molecule which, when transcribed in vivo, forms double-stranded RNA that induces the production of interferon. Expression cassettes also are provided comprising a promoter operably linked to a ribozyme or antisense nucleic acid molecule which, when transcribed in vivo, forms a ribozyme or antisense RNA molecule that stimulates an immune response. In addition, expression cassettes are provided comprising a promoter operably linked to a ribozyme or antisense nucleic acid molecule which, when transcribed in vivo, stimulates apoptosis. Finally, gene delivery vectors are provided which contain such expression cassettes, host cells containing the gene delivery vectors, and methods of utilizing the expression cassettes, gene delivery vectors, and host cells.

Abstract: The present invention relates techniques for identifying suitable secretion fusion partner (SFP) for hyper-secretory production of recombinant proteins. The SFPs can be obtained from secretome analyses. Recombinant proteins are produced in a fusion form with a secretion fusion partner (SFP) and can be separated from the SFP by in vitro protease treatment. SFPs of this invention greatly improve the secretion level of target proteins and peptides which are valuable for bio-pharmaceuticals and the bio-industry.

Abstract: Modified human interferon polypeptides and uses thereof are provided.

Abstract: Methods for high-throughput screening in duckweed are disclosed. In one aspect, these methods are used to identify nucleotide sequences encoding polypeptides of interest. In another aspect, these methods are used to identify nucleotide sequences that modulate the expression of a target nucleotide sequence. The methods combine the predictive benefits of screening in whole plants with the speed and efficiency of a high throughput system.

Abstract: Novel compositions for the production of interferons such as interferon-? 2a, interferon-? 2b, or interferon-? 1a (IFN-? 2a, IFN-? 2b, or IFN-? 1a) are provided. The compositions comprise components of vectors, such as a vector backbone, a promoter, and a gene of interest that encodes an interferon such as IFN-? 2a, IFN-? 2b, or IFN-? 1a, and the vectors comprising these components. In certain embodiments, these vectors are transposon-based vectors. Also provided are methods of making these compositions and methods of using these compositions for the production of an interferon such as IFN-? 2a, IFN-? 2b, or IFN-?1a.

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 has objects to provide a mutant interferon-? protein having a higher anti-viral and anti-tumor activity and to provide an agent for susceptive diseases, which contains the mutant interferon-? protein as an effective ingredient; and solves the above objects by providing a mutant protein which has an amino acid sequence of human interferon-? subtype ?8 represented by any one of SEQ ID NOs:1 to 3, where the arginine residue at the 145th has been replaced with leucine, isoleucine, or valine residue; the alanine residue at the 146th has been replaced with asparagine or serine residue; and the methionine residue at the 149th has been replaced with tyrosine residue; and an agent for susceptive diseases, containing the mutant protein.

Abstract: The present disclosure provides isolated Interferon? nucleic acids and polypeptides. The disclosure also provides antibodies which specifically recognize the subject Interferon? polypeptides, expression vectors containing the subject nucleic acids, and host cells expressing the subject polypeptides. In addition, methods of treatment using Interferon? are provided.

Abstract: Methods are disclosed for genetically engineering host cells that lack an endogenous pathway for fucosylating N-glycans of glycoproteins to be able to produce glycoproteins with fucosylated N-glycans.

Abstract: A method is described for the expression of heterologous polypeptides in rodent cells. The method comprises the oriP/EBNA-1 episomal replication and maintenance system of the Epstein-Barr-Virus (EBV). With the stable integration of an EBNA-1-protein expression cassette under the control of a promoter into the genome of a rodent cell an EBNA-1-protein expression in the cells was obtained. The heterologous protein is expressed from an episome comprising an EBV origin of replication and a functional expression cassette of said heterologous protein. The invention further comprises transformed rodent cell lines, a method for the production of a heterologous protein in said cell lines and a kit for the construction of said cell lines.

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: Human interferon-? protein analogs in which the asparagine at position 25, numbered in accordance with native human interferon-?, is recombinantly replaced with an aspartate residue exhibit a biological activity of human interferon-? (e.g. IFN-? 1b) at an increased level relative to IFN-? 1b. These analogs are obtained by introducing a gene coding for Asp25 IFN-? into a cell and expressing the recombinant protein. The resulting IFN-? protein analog is suitable for large scale manufacturing for incorporation in HA-containing or HA-free therapeutics for treatment of diseases including multiple sclerosis. A reduced Lys endoproteinase-C peptide map technique that produces a fingerprint profile for proteins using an enzymatic digest followed by RP-HPLC is also useful in quality control as an ID test for the IFN-? protein analog products.

Abstract: The invention provides compositions and methods of identifying, modifying and producing modified target molecules, including therapeutic molecules by modification with non-natural amino acids. Certain aspects of the invention include methods of adding a chemical moiety to a target molecule, and the compositions resulting therefrom. Certain aspects of the invention also relate to kits for identifying, modifying and producing modified target molecules described herein.

Abstract: The present invention is of a method of producing proteins in mammalian cells using a permanent selection in the absence of cytotoxic drugs. Specifically, the present invention can be used to produce large quantities of highly pure human proteins which are suitable for pharmaceutical applications.

Abstract: This application relates to recombinant human interferon-like proteins. In one embodiment a recombinant protein created by gene shuffling technology is described having enhanced anti-viral and anti-proliferative activities in comparison to naturally occurring human interferon alpha 2b (HuIFN-?2b). The invention encompasses a polynucleotide encoding the protein and recombinant vectors and host cells comprising the polynucleotide. Preferably the polynucleotide is selected from the group of polynucleotides each having a sequence at least 93% identical to SEQ ID: No. 1 and the protein is selected from the group of proteins each having an amino acid sequence at least 85% identical to SEQ ID No: 2. The proteins and compositions comprising the proteins can be used for treatment of conditions responsive to interferon therapy, such as viral diseases and cancer.

Abstract: A method for the production of high-level soluble human recombinant interferon alpha protein (rhuIFN?) in E. coli and vectors useful for such a production which comprises: (1) transforming an E. coli selected in the group consisting of E. coli protease deficient host strains, and E. coli reductase deficient host strains, with a recombinant expression vector comprising the sequence encoding the glutathione-S-transferase (GST), a junction sequence including a recognition site for a specific protease and a sequence able to encode an interferon alpha (IFN alpha) protein under the control of an inducible promoter, said vector encoding a GST-IFN alpha fusion protein; (2) expressing said interferon alpha protein in conditions comprising the induction of the expression with 0.1 mM-0.5 mM IPTG and a growth temperature of 25° and/or 37° C., depending on said E. coli strain; and (3) isolating the expressed IFN alpha protein.

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: The present disclosure provides an affinity polypeptide for the purification of a recombinant biologically active protein or polypeptide. Further, the present disclosure provides a fusion recombinant protein or polypeptide wherein the fusion recombinant protein comprises of at least two components, a biologically active polypeptide or protein or protein of interest and the affinity polypeptide. The biologically active polypeptide may be linked directly or indirectly to the affinity polypeptide by covalent binding. The present disclosure provides a recombinant expression vector for the producing said fusion recombinant protein in host cells. Further, the present disclosure provides an improved method of purification of recombinant protein from the host cells. Further, the disclosure provides a method of purification of the recombinant biologically active polypeptide or protein by immobilized metal ion chelating chromatography.

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 features mammalian expression systems with improved production yields, and method of using these systems to produce desired proteins. In one embodiment, the expression systems of the present invention comprise genetically-engineered mammalian host cells cultured in a medium that contains an effective amount of heparin or heparin-like molecules. The presence of heparin or heparin-like molecules significantly increases protein production by the cultured cells. The present invention also features the use of constitutively-active components of FGFR-I-mediated signal transduction pathways to improve protein production by cultured mammalian cells. Co-expression of such a component with a protein of interest markedly increases the production yield of the protein of interest.

Abstract: We describe a Cricetulus griseus cell which is modified, preferably genetically engineered, so that the expression of a heat shock protein is up-regulated compared to a cell which has not been so modified, as well as a method of expressing a recombinant protein from a host comprising such an engineered cell. Novel CHO heat shock protein sequences SEQ ID NO: 2 and SEQ ID NO: 4, as well as nucleic acid sequences SEQ ID NO: 1 and SEQ ID NO: 3 are also disclosed.

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: 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: A fusion polypeptide is described having the amino acid sequence X-Y-Z, or portion thereof, comprising the amino acid sequence of a glycosylated interferon-beta (X); Y is an optional linker moiety; and Z is a polypeptide comprising at least a portion of a polypeptide other than glycosylated interferon-beta. It is preferred that X is human interferon-beta-1a. Mutants of interferon-beta-1a are also described.

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 transfection under serum-free conditions. In preferred embodiments, the cells of use are SpESF or SpESF-X cells.

Abstract: The present invention relates to novel full-length interferon gamma (IFNG) polypeptide variants having interferon gamma activity. The full-length interferon gamma polypeptide variants of the invention are obtained by performing selected modifications in the C-terminal part of the molecule. The full-length interferon gamma polypeptide variants of the invention are useful in therapy, in particular for the treatment of interstitial pulmonary diseases, such as idiopathic pulmonary fibrosis.

Abstract: Disclosed are interferons of Rhesus and Cynomolgus origin and methods of production and use thereof.

Abstract: The present invention relates to biologically active polypeptides linked to one or more accessory polypeptides. 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: The present invention provides a method for increasing the yield of a protein produced by cultivating eukaryotic cells and adding an ionic substance to the culture medium prior to harvest of the protein. Suitable ionic substances are the salts of the Hofmeister series, amino acids and peptone.

Abstract: The present invention relates to polynucleotide and polypeptide molecules for Zcyto21, an interferon-like protein, which is most closely related to interferon-? at the amino acid sequence level. The present invention also includes antibodies to the Zcyto21 polypeptides, and methods of using the polynucleotides and polypeptides.

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: The present invention provides interferon-alpha polypeptides and conjugates, and nucleic acids encoding the polypeptides. The invention also includes compositions comprising these polypeptides, conjugates, and nucleic acids; cells containing or expressing the polypeptides, conjugates, and nucleic acids; methods of making the polypeptides, conjugates, and nucleic acids; and methods of using the polypeptides, conjugates, and nucleic acids.

Abstract: The present invention provides IFN-? conjugates including IFN-? peptides and modifying groups such as PEG moieties. The IFN-? peptide and modifying group are linked via an intact glycosyl linking group interposed between and covalently attached to the IFN-? peptide and the modifying group. The IFN-? conjugates are formed from glycosylated peptides by the action of a glycosyltransferase. The glycosyltransferase ligates a modified sugar onto an amino acid or a glycosyl residue on the IFN-? peptide. Also provided are methods for preparing the IFN-? conjugates, methods for treating various disease conditions with the IFN-? conjugates, and pharmaceutical formulations including the IFN-? conjugates.

Abstract: The invention relates to a conjugate exhibiting interferon ? (IFNB) activity and comprising at least one first non-polypeptide moiety covalently attached to an IFNB polypeptide, the amino acid sequence of which differs from that of wildtype human IFNB in at least one introduced and at least one removed amino acid residue comprising an attachment group for said first non-polypeptide moiety. The first non-polypeptide moiety is e.g. a polymer molecule or a sugar moiety. The conjugate finds particular use in therapy. The invention also relates to a glycosylated variant of a parent IFNB polypeptide comprising at least one in vivo glycosylation site, wherein an amino acid residue of said parent polypeptide located close to said glycosylation site has been modified to obtain the variant polypeptide having an increased glycosylation as compared to the glycosylation of the parent polypeptide.