Abstract: A G-CSF precursor comprising a signal peptide and a G-CSF peptide, wherein the signal peptide has the sequence of the human wild-type signal peptide of the human G-CSF/b molecule with at least one of the following mutations: deletion of Glu29, insertion of Glu26, substitution Lys11Leu, substitution His21Phe, and substitution Glu28Leu.

Abstract: Bi-specific fusion proteins with therapeutic uses are provided, as well as pharmaceutical compositions comprising such fusion proteins, and methods for using such fusion proteins to repair or regenerate damaged or diseased tissue. The bi-specific fusion proteins generally comprise: (a) a targeting polypeptide domain that binds to a target molecule; and (b) an activator domain that detectably modulates tissue regeneration.

Abstract: The present invention relates to granulocyte colony stimulating factor (G-CSF) modified with Y-shaped branched polyethylene glycol (YPEG-G-CSF) at a specific lysine (Lys 17) and the preparation thereof, as well as the pharmaceutical composition comprising YPEG-G-CSF and use thereof.

Abstract: A metal coordination complex of a biologically active moiety and a metal is disclosed. The complex confers to the biologically active moiety an improved performance which can include potency, stability, absorbability, targeted delivery, and combinations thereof.

Abstract: The present invention provides methods for the identification of CD4+ T-cell epitopes in the sequences of various proteins, namely, human cytokines and cytokine receptors, as well as the production of peptides which when incorporated into the protein sequence, are no longer capable of initiating the CD4+ T-cell response. In some embodiments, the present invention provides means and compositions suitable for reducing the immunogenicity of cytokines and cytokines receptors such as interferon-?, soluble tumor necrosis factor receptor-1, erythoropoietin, and thrombopoietin.

Abstract: Disclosed herein is pyrroline-carboxy-lysine (PCL), a pyrrolysine analogue, which is a natural, biosynthetically generated amino acid, and methods for biosynthetically generating PCL. Also disclosed herein are proteins, polypeptides and peptides that have PCL incorporated therein and methods for incorporating PCL into such proteins, polypeptides and peptides. Also disclosed herein is the site-specific derivatization of proteins, polypeptides and peptides having PCL or pyrrolysine incorporated therein. Also disclosed herein is the crosslinking of proteins, polypeptides and peptides having PCL or pyrrolysine incorporated therein.

Abstract: Conjugating LHRH to curcumin (LHRH-Curcumin) substantially enhances the bioavailability of curcumin, targets it to cells expressing LHRH receptors, facilitates intravenous administration, and increases the anti-cancer efficacy of curcumin. The conjugate may be used against cancer cells that express the LHRH receptor: pancreas, prostate, breast, testicular, uterine, ovarian, melanoma. LH-Curcumin conjugates may be used against cancer cells that express the LH receptor: prostate, breast, ovary, testis, uterus, pancreas, and melanoma.

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: 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: A process for high expression of protein of interest using an expression vector. The process comprises at least the following regulatory elements: a) a CMV promoter, or its functional variants, b) an intron, c) TPL or its functional variants, d) VA genes or functional variants, and e) a bovine growth hormone polyadenylation sequence or functional variants.

Abstract: The present invention relates to a fusion protein in which a carboxy terminal of human erythropoietin (EPO) is fused with a carboxy terminal peptide fragment of ? subunit of human chorionic gonadotropin (HCG), to DNA encoding the fusion protein, and to a method for preparation of the fusion protein. The fusion protein has the enhanced in vivo activity of erythropoietin.

Abstract: The invention provides nucleic acid molecules encoding FGF21 mutant polypeptides, FGF21 mutant polypeptides, pharmaceutical compositions comprising FGF21 mutant polypeptides, and methods for treating metabolic disorders using such nucleic acids, polypeptides, or pharmaceutical compositions.

Abstract: A complex comprising at least one target protein and at least one binding molecule having a binding affinity for said target protein, wherein said molecule having a binding affinity is covalently or non-covalently bound to at least one water-soluble polymer

Abstract: A complex of a polysaccharide and recombinant human BMP-2 and BMP-7, soluble at physiological pH, wherein the polysaccharide/BMP mass ratio is less than 15, the polysaccharide being selected from the group of polysaccharides having carboxyl functional groups, at least one of which is substituted with at least one hydrophobic radical.

Abstract: The present invention concerns methods and compositions for forming PEGylated complexes of defined stoichiometry and structure. In preferred embodiments, the PEGylated complex is formed using dock-and-lock technology, by attaching a target agent to a DDD sequence and attaching a PEG moiety to an AD sequence and allowing the DDD sequence to bind to the AD sequence in a 2:1 stoichiometry, to form PEGylated complexes with two target agents and one PEG moiety. In alternative embodiments, the target agent may be attached to the AD sequence and the PEG to the DDD sequence to form PEGylated complexes with two PEG moieties and one target agent. In more preferred embodiments, the target agent may comprise any peptide or protein of physiologic or therapeutic activity. The PEGylated complexes exhibit a significantly slower rate of clearance when injected into a subject and are of use for treatment of a wide variety of diseases.

Abstract: Methods for increasing and maintaining hematocrit in a mammal comprising administering a hyperglycosylated analog of erythropoietin are disclosed. An analog may be administered less frequently than an equivalent molar amount of recombinant human erythropoietin to obtain a comparable target hematocrit and treat anemia. Alternatively, a lower molar amount of a hyperglycosylated analog may be administered to obtain a comparable target hematocrit and treat anemia. Also disclosed are new hyperglycosylated erythopoietin analogs, methods of production of the analogs, and compositions comprising the analogs.

Abstract: The present invention relates to a method for preparing erythropoietin, wherein culture supernatant of erythropoietin-producing eukaryotic cells containing erythropoietin are subjected to the following steps: a) Removing the cell components; and b) treating the product from a) to the following chromatography steps in the sequence indicated i) reversed phase chromatography; ii) anion exchange chromatography; iii) hydroxyapatite chromatography.

Abstract: The present invention relates to a polymer comprising a cholane core having at least one derivatizable group covalently bonded thereto and a hydrophilic polymer chain covalently bonded to derivatizable group(s) and a process for producing it The present invention also relates to micellar aggregate formed from the polymer of the present.

Abstract: The invention provides, a nucleic acid molecule encoding a growth hormone (GH) in which the RSP sorting signal has been mutated, such that the GH can be constitutively secreted by the nonregulated secretory pathway (NRSP) in a mammalian cell. The invention also provides a nucleic acid molecule encoding a GH in which the three-dimensional conformation of the RSP sorting signal has been altered such that the GH can be constitutively secreted by the NRSP in a mammalian cell.

Abstract: FSH mutant with increased glycosylation and longer half-life is described. The use of this FSH mutant for inducing folliculogenesis in human patients is also described.

Abstract: A method for evaluating the cure level of a stroke patient comprises following steps: (1) obtaining isolated blood sample from said stroke patient; (2) determining the concentration of serum granulocyte colony-stimulating factor (G-CSF) of said blood sample; (3) comparing the relationship between said concentration of granulocyte colony-stimulating factor (G-CSF) and the stroke severity ranking of said stroke patient; wherein United State National Institute of Health Stroke Scale (NIHSS) or modified Ranking Scale (mRS) is used in said stroke severity ranking; and (4) Using said concentration of granulocyte colony-stimulating factor (G-CSF) to predict the possible cure level of said stroke patient. The invention further provides a prognosis biomarker for evaluating the cure level of a stroke patient, and a kit containing said prognosis biomarker.

Abstract: In certain aspects, the present invention provides compositions and methods for promoting bone growth and increasing bone density.

Abstract: A method of promoting wound healing or connective tissue reconstruction and a method of treating ischemia in a subject in need thereof are disclosed. The methods comprising topically administering to the subject about 10-30 mg per cm2 wound tissue of Erythropoietin and about 100-300 mg per cm2 wound tissue of Fibronectin, thereby promoting wound healing or connective tissue reconstruction or treating ischemia in the subject. Unit dosage forms, pharmaceutical compositions, cosmetic compositions and formulations comprising Erythropoietin and/or Fibronectin are also disclosed.

Abstract: The invention is a method for the purification of mono-PEGylated erythropoietin using two cation exchange chromatography steps wherein the same type of cation exchange material is used in both cation exchange chromatography steps and a method for producing a mono-PEGylated erythropoietin in substantially homogeneous form.

Abstract: The present invention relates to a method for the treatment and/or prophylaxis of multiple sclerosis, and to the use of erythropoietin for this purpose and for the manufacture of a medicament for the intermittent treatment and/or intermittent prophylaxis of multiple sclerosis.

Abstract: Disclosed is a method for production of human erythropoietin. By the method, the cells are cultured in a serum-free medium with repetitive medium exchanges, in which medium exchange is carried out either by collecting 80 to 95% of the culture when viable cell density has reached at 2×106˜4×106 cells/mL, or by adjusting the amount of the exchanged medium so that the initial density of the viable cells may be 1.5×105˜2.5×105 cells/mL.

Abstract: Methods of producing and purifying proteins that comprise a glycan tag are provided. In some embodiments, a protein may be modified to include a glycan tag so as to facilitate production of the protein, e.g., by promoting protein secretion and/or promoting protein solubility. In some embodiments, the present disclosure provides methods wherein a protein may be modified to include a glycan tag, which may then be used as an affinity tag for purification.

Abstract: The invention provides an efficient method of purification of a modified cytokine. The process includes the use of a chromatographic technique for the purification of the desired cytokine. The purified cytokine can be used as a therapeutic composition.

Abstract: This invention relates to a conjugate of a polymer moiety and an erythropoietin moiety.

Abstract: The present invention relates to mutants of human growth hormone, which contain newly introduced N-linked or O-linked glycosylation site(s), such that these recombinantly produced polypeptides have glycosylation patterns distinctly different from that of the naturally occurring human growth hormone. The polynucleotide coding sequences for the mutants, expression cassettes comprising the coding sequences, cells expressing the mutants, and methods for producing the mutants are also disclosed. Further disclosed are pharmaceutical compositions comprising the mutants and method for using the mutants.

Abstract: The invention relates to a modifying agent comprising a water soluble polymer, wherein the water soluble polymer comprises at least one reactive selenium group, said reactive selenium group being capable of reacting with a thiol group thereby forming an —Se—S— bond. Furthermore, the invention relates to a method for producing said modifying agents and their use in the modification of pharmaceutically active agents, e.g. G-CSF. Additionally, the invention concerns conjugates comprising a water-soluble polymer and a pharmaceutically active agent, wherein the water-soluble polymer is linked via a S—Se-bond to agent and a method for their production and their use as medicaments. Finally, the invention concerns a pharmaceutical composition comprising the inventive conjugates.

Abstract: The invention relates to the use of haematopoietic growth factors, in particular erythropoietin (EPO) and thrombopoietin (TPO), or derivatives, analogues or parts thereof, for promoting structural tissue regeneration.

Abstract: Conjugates between a protein and a water soluble polymer comprising multiple degradable carbonate linkages are provided.

Abstract: The current invention comprises a method for the regeneration of a cation exchange chromatography column.

Abstract: The present invention relates to methods for glycosylating a protein in which the protein is modified to include an alkyne and/or an azide group. The invention further relates to a protein glycosylated by these methods.

Abstract: The present invention relates to a method of producing a hydroxyalkyl starch derivative comprising reacting hydroxyalkyl starch of formula (I) at its reducing end which is not oxidized prior to said reaction, with a compound of formula (II) R?NH—R? (II) wherein R1-R2 and R3 are independently hydrogen or a linear or branched hydroxyalkyl group, and wherein either R? or R? or R? and R? comprise at least one functional group X capable of being reacted with at least one other compound prior to or after the reaction of (I) and (II), as well as to the hydroxyalkyl starch derivative as such, obtainable by said method, and to a pharmaceutical composition comprising said hydroxyalkyl starch derivative.

Abstract: A novel hemopoietin receptor gene (NR12) was successfully isolated by extracting motifs conserved among the amino acid sequences of known hemopoietin receptors and by using the predicted sequence. The NR12 gene encodes two forms of proteins, a transmembrane type and a soluble type. The expression of the NR12 gene was detected in tissues containing hematopoietic cells. NR12 is a novel hemopoietin receptor molecule involved in the regulation of immune system and hematopoiesis in vivo. Thus, NR12 is useful in the search for novel hematopoietic factors that functionally bind to the NR12 receptor, and in the development of therapeutic drugs for diseases associated with immunity or hematopoiesis.

Abstract: The present invention relates to novel endogenous variants of erythropoietin (EPO) and their use for treatment or prevention of a condition associated with tissue damage due to cell death (apoptosis, necrosis) and inflammation, in particular for neuroprotection, e.g. treatment of acute (for example stroke) and chronic disease (for example ALS) of the nervous system.

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

Abstract: Methods and materials are provided for the production of compositions of erythropoietin protein, wherein said compositions comprise a pre-selected N-linked glycosylation pattern as the predominant N-glycoform.

Abstract: The invention relates to a method for the preparation of a hydroxyalkyl starch derivative which comprises reacting hydroxyalkyl starch (HAS) via the optionally oxidised reducing end of the HAS with the amino group M of a crosslinking compound which, apart from the amino group, comprises a specifically protected carbonyl group, namely an acetal group or a ketal group.

Abstract: The invention includes a nucleic acid having a sequence at least 98% homologous to SEQ ID NO: 1, which encodes the ? subunit of canine thyroid stimulating hormone (TSH). The invention also includes a nucleic acid having a sequence at least 98% homologous to SEQ ID NO: 2, which encodes the ? subunit of canine TSH. The invention also includes a method of producing a recombinant canine thyroid stimulating hormone (rcTSH) subunit by expressing a nucleic acid having a sequence of SEQ ID NO: 1 and a nucleic acid having a sequence of SEQ ID NO: 2 in a transgenic insect cell modified to sialylate proteins and producing a sialylated rcTSH subunit. The insect cell may be a lepidopteran cell. The rcTSH may be used for diagnosis and treatment. It may be used to diagnose canine hypothyroidism.

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 of treating neurodegenerative disorders of the brain and spinal cord is disclosed. The therapeutic agent is a polyethylene glycol linked protein.

Abstract: The invention provides compositions, methods, and kits for increasing transport of a neurotrophin (e.g., erythropoietin (EPO)) across the blood brain barrier while allowing its activity to remain substantially intact. The neurotrophin (e.g., EPO) is transported across the blood brain barrier via one or more endogenous receptor-mediated transport systems.

Abstract: The use of human erythropoietin (EPO) to prevent or treat endothelial injury due to chemotherapy, radiation therapy, mechanical trauma, or to a disease state which damages the endothelium (such as inflammation, heart disease or cancer) is described. The use of EPO in conjunction with the administration of chemotherapeutic agents is described.

Abstract: When a pharmaceutical composition for preventing and/or treating peripheral neuropathy caused by chromosomal microtubule inhibition, which comprises erythropoietin as an active ingredient, is administered to patients receiving a microtubule inhibitor as an anticancer agent, such treatment alleviates peripheral neuropathy in the patients and enables increased dosage, prolonged period and increased frequency for administration of the above anticancer agent, which were never before achieved. Thus, the pharmaceutical composition not only contributes to improvement of QOL in the patients, but also enables prolongation of life in the patients.

Abstract: This invention provides methods for extracting platelets, compositions obtained therefore, and methods for using the same.

Abstract: A water soluble polymer comprising multiple degradable carbonate linkages in a backbone and, for each carbonate linkage in the backbone, an oligomer linked thereto by the carbonate linkage, wherein the oligomer is branched.

Abstract: The invention relates to erythropoietin liquid formulations that are stable in storage and to a method for the production thereof. The invention particularly relates to erythropoietin liquid formulations, which contain at least four amino acids selected from the group consisting of leucine, isoleucine, threonine, glutamic acid, aspartic acid and phenylalanine, and in which the addition of preservatives, urea or human serum albumine can be foregone.