Abstract: Novel polypeptide compositions based on the amino acid sequence of tissue plasminogen activator (tPA) are provided having improved properties over natural tissue plasminogen activator. Particularly, enhanced specific activity, reduced response to inhibition by plasminogen activator inhibitor, fibrin stimulation of plasminogenolytic activity and/or enhanced affinity to fibrin surfaces are provided by modifying one or more loci by deletions or substitutions. One or both of the N- or C-termini may be modified.

Abstract: Novel polypeptide compositions based on the amino acid sequence of tissue plasminogen activator (tPA) are provided having improved properties over natural tissue plasminogen activator. Particularly, enhanced specific activity, reduced response to inhibition by plasminogen activator inhibitor, fibrin stimulation of plasminogenolytic activity and/or enhanced affinity to fibrin surfaces are provided by modifying one or more loci by deletions or substitutions. One or both of the N- or C-termini may be modified.The plasmids designated 1, 2 and 3 were deposited at the A.T.C.C. on Dec. 20, 1985 and given A.T.C.C. designations 40214, 40215 and 40216, respectively.

Abstract: A method for refolding recombinant platelet-derived growth factor from a high expression host cell system, such as E. coli, is provided in accordance with the present invention. The recombinant platelet-derived growth factor is isolated from inclusion body proteins, after which the free sulfydryl groups of the reduced monomeric recombinant protein are blocked. The blocked recombinant protein can then be brought into a biologically active conformation, without interference from the formation of incorrect disulfide bonds. Once in a biologically active conformation, the recombinant platelet-derived growth factor may be unblocked to allow the desired disulfide bonds to form, thus locking the recombinant protein into a biologically active conformation. The present invention also provides a novel mixed disulfide intermediate wherein the free sulfhydryl groups of reduced recombinant platelet-derived growth factor are derivatized to a disulfide blocking agent.

Abstract: Mammalian granulocyte-colony stimulating factor (G-CSF) receptor proteins, DNAs and expression vectors encoding mammalian G-CSF receptors, and processes for producing mammalian G-CSF receptors as products of recombinant cell culture, are disclosed.

Abstract: Thrombolytic proteins are disclosed which have tissue plasminogen-type activity. The proteins are characterized by modification within the 94 amino acid N-terminus, and/or at Arg-275, and/or at one or more of the N-linked glycosylation sites. Methods for making these proteins are disclosed as are therapeutic compositions containing same.

Abstract: Tissue plasminogen activator (t-PA) zymogens and variants are prepared, including a fibrinolytically active variant of t-PA that has an amino acid alteration at a site within the protease domain of t-PA as compared with the corresponding wild-type t-PA, which alteration renders the variant zymogenic in the presence of plasmin-degraded fibrinogen, and/or fibrin (or plasma clot) specific, as compared to the corresponding wild-type t-PA. DNA sequences can be prepared that encode the zymogens and variants, as well as expression vectors incorporating the DNA sequences, and host cells transformed with the expression vectors. The zymogens and variants may be used in a pharmaceutical preparation to treat a vascular disease or condition or to prevent fibrin deposition or adhesion formation or reformation in mammals.

Abstract: A method for refolding insoluble, improperly folded IGF-I is provided, wherein the IGF-I, precipitated from prokaryotic host cells, is concurrently solubilized, unfolded, and refolded into a biologically active conformation in a single buffer. The buffer contains reducing agent and chaotropic agent to solubilize the IGF-I at concentrations sufficiently low to allow solubilization and refolding to occur. Also provided is a triple-protease deficient E. coli host suitable for use in the process.

Abstract: A TPA variant with replacement of amino acids is provided. In a typical embodiment, the variant has an amino acid sequence with replacement of the amino acid group consisting of asparagine at the 37th position, serine at the 38th position, glycine at the 39th position, arginine at the 40th position, alanine at the 41st position, and glutamine at the 42nd position from the N-terminus of the amino acid sequence of mature human tissue plasminogen activator.

Abstract: The present invention relates to the development of bone growth factors as therapeutics for the prevention and treatment of pathological conditions involving bone tissue. The present invention provides biologically active proteinaceous factors comprising polypeptides containing the amino acid sequence M-A-G-L-G-D-E-F-G-D, [SEQ ID NO: 1], (X) F E T L F G A/V E D V I/D A L Q F V C G D, [SEQ ID NO: 2], or A-Y-I-P-I-E-T-L-E-G-I/G-E-L-V-D/Q-T-G/L-Q-F, [SEQ ID NO: 3], or biologically active fragments or sequence analogues thereof. Among the biological properties of the proteinaceous materials of the present invention is the capability to promote the growth and/or differentiation of osteoblastic cells.

Abstract: Tissue plasminogen activator (t-PA) zymogens and variants are prepared, including a fibrinolytically active variant of t-PA that has an amino acid alteration at a site within the protease domain of t-PA as compared with the corresponding wild-type t-PA, which alteration renders the variant zymogenic in the presence of plasmin-degraded fibrinogen, and/or fibrin (or plasma clot) specific, as compared to the corresponding wild-type t-PA. DNA sequences can be prepared that encode the zymogens and variants, as well as expression vectors incorporating the DNA sequences, and host cells transformed with the expression vectors. The zymogens and variants may be used in a pharmaceutical preparation to treat a vascular disease or condition or to prevent fibrin deposition or adhesion formation or reformation in mammals.

Abstract: A simple and efficient method of preparing active recombinant human neutrophil chemotactic factor. The human neutrophil chemotactic factor (NCF) is solubilized from a transformant cell homogenate using urea or guanidine hydrochloride. The polypeptide is further purified from the solution using conventional protocols such as CM-Sepharose CL-6B, HW-55 column, etc.. By using this method, large amounts of active human NCF can be recovered and purified.

Abstract: DNA molecules encoding mutant forms of bone morphogenetic proteins (BMP) are disclosed, The mutant forms of BMP can be produced bacterially and refolded to produce biologically active homodimers or heterodimers of BMP. A method of making such mutant BMPs is also disclosed.

Abstract: An expression vector comprises a cDNA sequence encoding an aFGF protein and a T7 promoter upstream therefrom. The vector is useful for transforming host cells and expressing the aFGF gene.

Abstract: The present invention relates to novel fragments of basic fibroblast growth factor (bFGF). The mitogenic potency of one of these bFGF fragments, identified as HBF-2 is about 25-50 fold less than that of native bFGF but at least 10.sup.3 -10.sup.4 fold more active than that of previously reported synthetic fragments of bFGF. Therefore, the present invention provides the shortest fragment of bFGF that retains substantial biologic activity.

Abstract: A fibrinolytically active amino acid sequence variant of a plasminogen activator is prepared that has one or more glycosylation sites in regions that are not glycosylated in the native molecule. DNA sequences can be prepared that encode the variants, as well as expression vectors incorporating the DNA sequences, and host cells transformed with the expression vectors. The variants may be used in a pharmaceutical preparation to treat a vascular disease or condition in patients.

Abstract: The present invention provides a method for increasing expression of low molecular weight polypeptides which are produced by recombinant DNA tranformation of prokaryotic host cells. This invention allows the direct expression of these polypeptides without fusion to an otherwise expressible protein and requires only the addition of a single amino acid to the sequence of the polypeptide product of interest.

Abstract: Des-epidermal growth factor homologous--plasminogen activators with greatly diminished affinity for liver membranes.

Abstract: A novel polypeptide, designated neurotrophic factor-4 (NT-4), has been identified by PCR amplification of human genomic DNA. Provided herein is nucleic acid encoding NT-4 useful in diagnostics and in the recombinant preparation of NT-4. NT-4 is used in the treatment of nerve cells and in diagnostic assays.

Abstract: This invention provides an isolated nucleic acid molecule encoding a human 5-HT.sub.1F receptor, an isolated protein which is a human 5-HT.sub.1F receptor, vectors comprising an isolated nucleic acid molecule encoding a human 5-HT.sub.1F receptors, mammalian cells comprising such vectors, antibodies directed to the human 5-HT.sub.1F receptor, nucleic acid probes useful for detecting nucleic acid encoding human 5-HT.sub.1F receptors, antisense oligonucleotides complementary to any sequences of a nucleic acid molecule which encodes a human 5-HT.sub.1F receptor, pharmaceutical compounds related to human 5-HT.sub.1F receptors, and nonhuman transgenic animals which express DNA a normal or a mutant human 5-HT.sub.1F receptor. This invention further provides methods for determining ligand binding, detecting expression, drug screening, and treatment involving the human 5-HT.sub.1F receptor.

Abstract: An analog of porcine growth hormone, is disclosed which retains the diabetogenic, insulin-sparing and lipolytic properties of porcine growth hormone while being capable of improving growth in mammals.